Methods of treating oral mucositis

ABSTRACT

Methods and kits for treating oral mucositis are disclosed. The treatment comprises administering to a patient in need thereof a Reactive Oxygen Species scavenger in a pharmaceutically acceptable formulation.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority from U.S. patent application Ser. No. 15/841,586 filed on Dec. 14, 2017, now U.S. Pat. No. 10,610,533, which claims priority from U.S. patent application Ser. No. 11/871,848 filed on Oct. 12, 2007, now U.S. Pat. No. 9,855,279, which claims priority from U.S. Provisional Application Ser. No. 60/829,291 filed Oct. 12, 2006, each of which are incorporated herein by reference.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

Not Applicable.

INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ON A COMPACT DISC

Not Applicable.

BACKGROUND OF THE INVENTION 1. Field of the Invention

This invention relates generally to the treatment of oral mucositis and, more particularly, to methods for treating oral mucositis with a Reactive Oxygen Species (“ROS”) scavenger. The compositions and methods are useful in treating oral mucositis.

2. Description of Related Art

Oral ulcerative mucositis is a common, painful, dose-limiting toxicity of drug and radiation therapy for cancer (1). The disorder is characterized by breakdown of the oral mucosa that results in the formation of ulcerative lesions. In granulocytopenic patients, the ulcerations that accompany mucositis are frequent portals of entry for indigenous oral bacteria often leading to sepsis or bacteremia (2). Mucositis occurs to some degree in more than one third of patients receiving anti-neoplastic drug therapy (3). The frequency and severity are significantly greater among patients who are treated with induction therapy for leukemia or with many of the conditioning regimens for bone marrow transplant (4). Among these individuals, moderate to severe mucositis can occur in more than three-quarters of patients. Moderate to severe mucositis occurs in virtually all patients who receive radiation therapy for tumors of the head and neck and typically begins with cumulative exposures of 15 Gy and then worsens as total doses of 60 Gy or more are reached (1-4).

Clinically mucositis progresses through three stages:

1. Atrophic changes accompanied by painful mucosal erythema, which can respond to local anesthetics.

2. Painful ulceration with pseudomembrane formation and, in the case of myelosuppressive treatment, potentially life-threatening sepsis, requiring antimicrobial therapy. Pain is often of such intensity as to require parenteral narcotic analgesia.

3. Spontaneous healing, occurring about 2-3 weeks after cessation of anti-neoplastic therapy.

Standard therapy for mucositis is predominantly palliative, including application of topical analgesics such as lidocaine and/or systemic administration of narcotics and antibiotics. At present, the only approved treatment for oral mucositis is palifermin (Kepivance) which is a member of the fibroblast growth factor (FGF) superfamily of molecules. Palifermin's approval is limited to the treatment of oral mucositis in the patients undergoing conditioning regimens prior to hematopoietic stem cell transplants for the treatment of hematologic malignancies.

The complexity of mucositis as a biological process has only been recently appreciated (5-7). It has been suggested that the condition represents a sequential interaction of oral mucosal cells and tissues, reactive oxygen species, pro-inflammatory cytokines, mediators of apoptosis and local factors such as saliva and the oral microbiota. While epithelial degeneration and breakdown ultimately result in mucosal ulceration, it appears that the early changes associated with radiation-induced mucosal toxicity occur within the endothelium, and connective tissue of the submucosa. For example, electron microscopic evaluation of mucosa within 1 week of radiation shows damage to both endothelium and connective tissue, but not epithelium. It appears that the overall mechanism for mucositis development is similar for both radiation and chemotherapy (8).

Recently, a superoxide dismutase mimetic, M40403, was shown to be effective in an animal models of inflammation (Salvemini et al., Science 286:304-306, 1999) and, more specifically, in an animal model of rheumatoid arthritis (Salvemini et al., Arthritis & Reumatism 44:2909-2921, 2001). Nevertheless, treatment of oral mucositis using M40403 has neither been reported nor suggested.

BRIEF SUMMARY OF THE INVENTION

The present teachings provide methods for treating oral mucositis. A method comprises administering to a patient in need thereof, a superoxide dismutase mimetic. In accordance with one aspect of the invention, the superoxide dismutase mimetic can be represented by the formula:

wherein

(i) R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀ are independently:

(i^(a)) hydrogen; or

(i) a moiety independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, heterocyclyl, and aralkyl radicals and radicals attached to the α-carbon or α-amino acids; or

(i^(c)) a moiety independently selected from the group consisting of —OR₁₁, —NR₁₁R₁₂, —COR₁₁, —CO₂R₁₁, —CONR₁₁R₁₂, —SR₁₁, —SOR₁₁, —SO₂R₁₁, —SO₂NR₁₁R₁₂, —N(OR₁₁)(R₁₂), —P(O)(OR₁₁)(OR₁₂), —P(O)(OR₁₁)(R₁₂), —OP(O)(OR₁₁)(OR₁₂), and substituents attached to the α-carbon of α-amino acids, wherein R₁₁ and R₁₂ are independently hydrogen or alkyl; and

(ii) optionally, one or more of R₁ or R′₁ and R₂ or R′₂, R₃ or R′₃ and R₄ or R′₄, R₅ or R′₅ and R₆ or R′₆, R₇ or R′₇ and R₈ or R′₈, R₉ or R′₉ and R₁₀ or R′₁₀ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(iii) optionally, one or more of R₁ and R′₁, R₂ and R′₂, R₃ and R′₃, R₄ and R′₄, R₅ and R′₅, R₆ and R′₆, R₇ and R′₇, R₈ and R′₈, R₉ and R′₉, and R₁₀ and R′₁₀, together with the carbon atom to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(iv) optionally, one or more of R₁₀ or R′₁₀ and R₁ or R′₁, R₂ or R′₂ and R₃ or R′₃, R₄ or R′₄ and R₅ or R′₅, R₆ or R′₆ and R₇ or R′₇, or R₈ or R′₈ and R₉ or R′₉ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted nitrogen containing heterocycle having 3 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(v) optionally, one or more of R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀, together with a different one of R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀, which is attached to a different carbon atom in the macrocyclic ligand may be bound to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(vi) combinations of any of (i) through (v) above;

wherein

M is a transition metal;

X, Y and Z are independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof; or

X, Y and Z are independently selected from the group consisting of charge-neutralizing anions which are derived from any monodentate or polydentate coordinating ligand and a ligand system and the corresponding anion thereof; or

X, Y and Z are independently attached to one or more of R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀; and

n is an integer from 0 to 3.

Preferably, M is selected from the group consisting of Mn²⁺, Mn³⁺, Mn⁴⁺, Mn⁶⁺, Mn⁷⁺, Fe²⁺, Fe³⁺, Fe⁴⁺, Fe⁵⁺, Ni²⁺, Ni³⁺, Cu¹⁺, Cu²⁺, V²⁺, V³⁺, V⁴⁺, and V⁵⁺.

In an alternative, the superoxide dismutase mimetic can be represented by the formula:

wherein

(i) a nitrogen of the macrocycle and two adjacent carbon atoms to which the nitrogen is attached independently form a substituted or unsubstituted, saturated, partially saturated or unsaturated nitrogen-containing heterocycle W having 2 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(ii) one or more of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀ are independently:

(ii^(a)) hydrogen; or

(ii^(b)) a moiety independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, heterocyclyl, and aralkyl radicals and radicals attached to the α-carbon or α-amino acids; or

(ii^(c)) a moiety independently selected from the group consisting of —OR₁₁, —NR₁₁R₁₂, —COR₁₁, —CO₂R₁₁, —CONR₁₁R₁₂, —SR₁₁, —SOR₁₁, —SO₂R₁₁, —SO₂NR₁₁R₁₂, —N(OR₁₁)(R₁₂), —P(O)(OR₁₁)(OR₁₂), —P(O)(OR₁₁)(R₁₂), —OP(O)(OR₁₁)(OR₁₂), and substituents attached to the α-carbon of α-amino acids, wherein R₁, and R₁₂ are independently hydrogen or alkyl; and

(iii) optionally, one or more of R₁ and R₂ or R′₂, R₃ or R′₃ and R₄ or R′₄, R₅ or R′₅ and R₆ or R′₆, R₇ or R′₇ and R₈ or R′₈, R₉ or R′₉ and R₁₀ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(iv) optionally, one or more of R₂ and R′₂, R₃ and R′₃, R₄ and R′₄, R₅ and R′₅, R₆ and R′₆, R₇ and R′₇, R₈ and R′₈, and R₉ and R′₉, together with the carbon atom to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(v) optionally, one or more of R₂ or R′₂ and R₃ or R′₃, R₄ or R′₄ and R₅ or R′₅, R₆ or R′₆ and R₇ or R′₇, or R₈ or R′₈ and R₉ or R′₉ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted nitrogen containing heterocycle having 3 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(vi) optionally, one or more of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀, together with a different one of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀, which is attached to a different carbon atom in the macrocyclic ligand may be bound to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(vii) optionally, one or more of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀, may be bound to an atom of heterocycle W to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(viii) combinations of any of (i) through (vii) above;

wherein

M is a transition metal;

X, Y and Z are independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof; or

X, Y and Z are independently selected from the group consisting of charge-neutralizing anions which are derived from any monodentate or polydentate coordinating ligand and a ligand system and the corresponding anion thereof; or

X, Y and Z are independently attached to one or more of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀; and

n is an integer from 0 to 3.

Preferably, M is selected from the group consisting of Mn²⁺, Mn³⁺, Mn⁴⁺, Mn⁶⁺, Mn⁷⁺, Fe²⁺, Fe³⁺, Fe⁴⁺, Fe⁶⁺, Ni²⁺, Ni³⁺, Cu¹⁺, Cu²⁺, V²⁺, V³⁺, V⁴⁺, and V⁵⁺, and W is a substituted or unsubstituted pyridino moiety.

In yet another alternative, the superoxide dismutase mimetic can be represented by the formula:

wherein

(i) a nitrogen of the macrocycle and two adjacent carbon atoms to which the nitrogen is attached independently form a substituted or unsubstituted, saturated, partially saturated or unsaturated nitrogen-containing heterocycle W having 2 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(ii) two sets of two adjacent carbon atoms of the macrocycle independently form substituted or unsubstituted, saturated, partially saturated or unsaturated, cycles or heterocycles U and V having 3 to 20 carbon atoms; and

(iii) R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀ are independently:

(iii^(a)) hydrogen; or

(iii^(b)) a moiety independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, heterocyclyl, and aralkyl radicals and radicals attached to the α-carbon or α-amino acids; or

(iii^(c)) a moiety independently selected from the group consisting of —OR₁₁, —NR₁₁R₁₂, —COR₁₁, —CO₂R₁₁, —CONR₁₁R₁₂, —SR₁₁, —SOR₁₁, —SO₂R₁₁, —SO₂NR₁₁R₁₂, —N(OR₁₁)(R₁₂), —P(O)(OR₁₁)(OR₁₂), —P(O)(OR₁₁)(R₁₂), —OP(O)(OR₁₁)(OR₁₂), and substituents attached to the α-carbon of α-amino acids, wherein R₁₁ and R₁₂ are independently hydrogen or alkyl; and

(iv) optionally, one or more of R₁ and R₂ or R′₂, R₅ or R′₅ and R₆ or R′₆, R₉ or R′₉ and R₁₀ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(v) optionally, one or more of R₂ and R′₂, R₅ and R′₅, R₆ and R′₆, and R₉ and R′₉, together with the carbon atom to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(vi) optionally, one or more of R₂ or R′₂ and R₃, R₄ and R₅ or R′₅, R₆ or R′₆ and R₇, or R₈ and R₉ or R′₉ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted nitrogen containing heterocycle having 3 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(vii) optionally, one or more of R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₆, R₉, R′₉, and R₁₀, together with a different one of R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, Ra, R₉, R′₉, and R₁₀, which is attached to a different carbon atom in the macrocyclic ligand may be bound to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(viii) optionally, one or more of R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀, may be individually bound to an atom of heterocycles U, V and W to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(ix) combinations of any of (i) through (viii) above;

wherein

M is a transition metal;

X, Y and Z are independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof; or

X, Y and Z are independently selected from the group consisting of charge-neutralizing anions which are derived from any monodentate or polydentate coordinating ligand and a ligand system and the corresponding anion thereof; or

X, Y and Z are independently attached to one or more of R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀; and

n is an integer from 0 to 3.

Preferably, M is selected from the group consisting of Mn²⁺, Mn³⁺, Mn⁴⁺, Mn⁶⁺, Mn⁷⁺, Fe²⁺, Fe³⁺, Fe⁴⁺, Fe⁶⁺, Ni²⁺, Ni³⁺, Cu¹⁺, Cu²⁺, V²⁺, V³⁺, V⁴⁺, and V⁵⁺. In accordance with a futher aspect of the invention, U and V are saturated cycloalkyl heterocycles having 3 to 20 carbon atoms preferably saturated cycloalkyl heterocycles having 4 to 10 carbon atoms, and still more preferably U and V are trans-cyclohexanyl fused rings. In yet another aspect of the present invention, W is a substituted or unsubstituted pyridino moiety, more preferably, U and V are trans-cyclohexanyl fused rings and W is a substituted pyridino moiety. Preferably, the superoxide dismutase mimetic can be represented by the formula:

The ROS scavenger can be administered in an amount of at most 0.015 mg/kg, or preferably at most 2 mg/kg. In yet another aspect, the pharmaceutically acceptable formulation is a pharmaceutically acceptable oral formulation and administering comprises administering orally. Preferably, the patient is a human patient and the oral mucositis is a result of chemotherapy or radiation therapy.

Another aspect of the invention can be a method of treating a cancer, the method comprising: a) administering to a subject in need of cancer treatment a pharmaceutical composition comprising a superoxide dismutase mimetic; and b) administering to the subject an effective amount of a cancer treatment, whereby the superoxide dismutase mimetic prevents or reduces oral mucositis in the subject.

The cancer treatment can be comprised of radiation therapy and chemotherapy. In said method of treating a cancer, the superoxide dismutase mimetic can be a reactive oxygen species scavenger, and the pharmaceutical composition can further comprise at least one additional reactive oxygen species scavenger selected from the group consisting of amifostine and N-acetylcysteine. Additionally, the method of treating a cancer can further comprise administering a pharmaceutical composition which upregulates expression of at least one transcription factor which increases expression of one or more genes controlling at least one naturally occurring antioxidant pathway. The composition which upregulates expression of at least one transcription factor can be palifermin, and the transcription factor upregulated can be Nrf-2.

In yet another aspect of the present invention, a kit can be provided for treating oral mucositis, the kit comprising: (a) a ROS scavenger, wherein the ROS scavenger can be a superoxide dismutase; (b) one additional pharmaceutical compound selected from the group consisting of a chemotherapeutic agent; (c) a non-superoxide dismutase mimetic radical scavenger; and (d) instructions for administering the superoxide dismutase to a subject in need of cancer therapy. A chemotherapeutic agent can be selected from a group consisting of all-trans retinoic acid, azacitidine, azathioprine, bleomycin, carboplatin, capecitabine, cisplatin, chlorambucil, cyclophosphamide, cytarabine, daunorubicin, docetaxel, doxifluridine, doxorubicin, epirubicin, epothilone, etoposide, fluorouracil, gemcitabine, hydroxyurea, idarubicin, imatinib, mechlorethamine, mercaptopurine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, teniposide, tiguanine, valrubicin, vinblastine, vincristine, vindesine, vinorelbine. A non-superoxide dismutase mimetic radical scavenger can be selected from the group of amifostine and N-acetylcysteine. A superoxide dismutase mimetic can be M40403.

These and other features, aspects and advantages of the present invention will become better understood with reference to the following description, examples and appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a timeline of a study for treatment groups 1-6 to evaluate the effect of M40403, administered by either ip or topical routes. The grade of mucositis was scored, beginning on day 6, and for every second day thereafter, through and including day 28.

FIG. 2 is a validated photographic scale for mucositis scoring.

FIG. 3 illustrates the percent daily weight change for the groups treated with intraperitoneal (ip) injection of M40403 (i.e. groups 1, 2, 3, and 6), including the control group, and for the groups treated topically with M40403 (i.e. groups 4 and 5).

FIG. 4 illustrates the mean percent weight gain for the groups treated ip with M40403 (i.e. groups 1, 2, 3, and 6), including the control group, and for the groups treated topically with M40403 (i.e. groups 4 and 5).

FIG. 5 illustrates the mean daily mucositis scores the groups treated ip with M40403 (i.e. groups 1, 2, 3, and 6), including the control group, and for the groups treated topically with M40403 (i.e. groups 4 and 5).

FIG. 6 illustrates the percentage of animal days with a mucositis score of 3 or higher for groups treated ip with M40403 (i.e. groups 1, 2, 3, and 6), including the control group, and for the groups treated topically with M40403 (i.e. groups 4 and 5).

FIG. 7 is a timeline of a study for treatment groups 1-9 to evaluate the effect of M40403, administered by either ip or topical routes. The grade of mucositis was scored, beginning on day 6, and for every second day thereafter, through and including day 28.

FIG. 8 is a validated photographic scale for mucositis scoring.

FIG. 9 illustrates the percent daily weight change for the groups treated ip with M40403 (i.e. groups 2-9) and for the control group (i.e. group 1).

FIG. 10 illustrates the mean percent weight gain for the groups treated ip with M40403 (i.e. groups 2-9) and for the control group (i.e. group 1).

FIG. 11 illustrates the mean daily mucositis scores the groups treated ip with M40403 (i.e. groups 2-9) and for the control group (i.e. group 1).

FIG. 12 illustrates the percentage of animal days with a mucositis score of 3 or higher for groups treated ip with M40403 (i.e. groups 2-9) and for the control group (i.e. group 1).

DETAILED DESCRIPTION OF THE INVENTION

Abbreviations and Definitions

To facilitate understanding of the invention, a number of terms and abbreviations as used herein are defined below as follows:

The term “alkenyl”, alone or in combination, means an alkyl substituent having one or more double bonds. Examples of such alkenyl substituents include, but are not limited to, ethenyl, propenyl, 1-butenyl, cis-2-butenyl, trans-2-butenyl, iso-butylenyl, cis-2-pentenyl, trans-2-pentenyl, 3-methyl-1-butenyl, 2,3-dimethyl-2-butenyl, 1-pentenyl, 1-hexenyl, 1-octenyl, decenyl, dodecenyl, tetradecenyl, hexadecenyl, cis- and trans-9-octadecenyl, 1,3-pentadienyl, 2,4-pentadienyl, 2,3-pentadienyl, 1,3-hexadienyl, 2,4-hexadienyl, 5,8,11,14-eicosatetraenyl, and 9,12,15-octadecatrienyl.

The term “alkyl”, alone or in combination, means a straight-chain or branched-chain alkyl substituent containing from 1 to about 22 carbon atoms, preferably from about 1 to about 18 carbon atoms, and most preferably from about 1 to about 12 carbon atoms. Examples of such substituents include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl and eicosyl.

The terms “alkylcycloalkyl” and “alkenylcycloalkyl” mean a cycloalkyl substituent as defined above which is substituted by an alkyl or alkenyl substituent as defined above. Examples of alkylcycloalkyl and alkenylcycloalkyl substituents include, but are not limited to, 2-ethylcyclobutyl, 1-methylcyclopentyl, 1-hexylcyclopentyl, 1-methylcyclohexyl, 1-(9-octadecenyl)cyclopentyl and 1-(9-octadecenyl)cyclohexyl.

The terms “alkylcycloalkenyl” and “alkenylcycloalkenyl” means a cycloalkenyl substituent as defined above which is substituted by an alkyl or alkenyl substituent as defined above. Examples of alkylcycloalkenyl and alkenylcycloalkenyl substituents include, but are not limited to, 1-methyl-2-cyclopentyl, 1-hexyl-2-cyclopentenyl, 1-ethyl-2-cyclohexenyl, 1-butyl-2-cyclohexenyl, 1-(9-octadecenyl)-2-cyclohexenyl and 1-(2-pentenyl)-2-cyclohexenyl.

The term “alkynyl”, alone or in combination, means an alkyl substituent having one or more triple bonds. Examples of such alkynyl groups include, but are not limited to, ethynyl, propynyl (propargyl), 1-butynyl, 1-octynyl, 9-octadecynyl, 1,3-pentadiynyl, 2,4-pentadiynyl, 1,3-hexadiynyl, and 2,4-hexadiynyl.

The term “aralkyl”, alone or in combination, means an alkyl or cycloalkyl substituent as defined above in which one hydrogen atom is replaced by an aryl substituent as defined above, such as benzyl, 2-phenylethyl, and the like.

The term “aryl”, alone or in combination, means a phenyl or naphthyl substituent which optionally carries one or more substituents selected from alkyl, cycloalkyl, cycloalkenyl, aryl, heterocycle, alkoxyaryl, alkaryl, alkoxy, halogen, hydroxy, amine, cyano, nitro, alkylthio, phenoxy, ether, trifluoromethyl and the like, such as phenyl, p-tolyl, 4-methoxyphenyl, 4-(tert-butoxy)phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-hydroxyphenyl, 1-naphthyl, 2-naphthyl, and the like.

The term “cycloalkenyl”, alone or in combination, means a cycloalkyl substituent having one or more double bonds. Examples of cycloalkenyl substituents include, but are not limited to, cyclopentenyl, cyclohexenyl, cyclooctenyl, cyclopentadienyl, cyclohexadienyl and cyclooctadienyl.

The terms “cyclic”, “cycle” or “cycylyl” means a ring structure containing 3 to 20 carbon atoms, preferably 5 to 10 carbon atoms, which may be heterocyclic. The cyclic, cycle or cycylyl can also contain more than one ring.

The term “cycloalkenylalkyl” means an alkyl substituent as defined above which is substituted by a cycloalkenyl substituent as defined above. Examples of cycloalkenylalkyl substituents include, but are not limited to, 2-cyclohexen-1-ylmethyl, 1-cyclopenten-1-ylmethyl, 2-(1-cyclohexen-1-yl)ethyl, 3-(1-cyclopenten-1-yl)propyl, 1-(1-cyclohexen-1-ylmethyl)pentyl, 1-(1-cyclopenten-1-yl)hexyl, 6-(1-cyclohexen-1-1-yl)hexyl, 1-(1-cyclopenten-1-yl)nonyl and 1-(1-cyclohexen-1-yl)nonyl.

The term “cycloalkyl”, alone or in combination means a cycloalkyl radical containing from 3 to about 10, preferably from 3 to about 8, and most preferably from 3 to about 6, carbon atoms. Examples of such cycloalkyl substituents include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and perhydronaphthyl.

The term “cycloalkylalkyl” means an alkyl substituent as defined above which is substituted by a cycloalkyl substituent as defined above. Examples of cycloalkylalkyl substituents include, but are not limited to, cyclohexylmethyl, cyclopentylmethyl, (4-isopropylcyclohexyl)methyl, (4-t-butyl-cyclohexyl)methyl, 3-cyclohexylpropyl, 2-cyclohexylmethylpentyl, 3-cyclopentylmethylhexyl, 1-(4-neopentylcyclohexyl)methylhexyl, and 1-(4-isopropylcyclohexyl)methylheptyl.

The term “cycloalkylcycloalkyl” means a cycloalkyl substituent as defined above which is substituted by another cycloalkyl substituent as defined above. Examples of cycloalkylcycloalkyl substituents include, but are not limited to, cyclohexylcyclopentyl and cyclohexylcyclohexyl.

The term “halide” means chloride, fluoride, iodide, or bromide.

The term “heterocyclic”, “heterocycle” or “heterocycylyl” means a cyclic, cycle or cycylyl containing at least one other kind of atom, in addition to carbon, in the ring. Such atoms include, but are not limited to, nitrogen, oxygen and sulfur. The heterocyclic can also contain more than one ring. Examples of heterocyclics include, but are not limited to, pyrrolidinyl, piperidyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl, thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl, benzothiadiazolyl, triazolyl and tetrazolyl groups.

The term “nitrogen containing heterocycle” means a ring structure in which 2 carbons and a nitrogen of the ring are shared with the fifteen-membered macrocyclic ligand. The nitrogen containing heterocycle can contain 2 to 20, preferably 4 to 10, carbon atoms, can be substituted or unsubstituted, saturated, partially saturated or unsaturated, and can also contain nitrogen, oxygen and/or sulfur atoms in the portion of the ring which is not also part of the fifteen-membered macrocyclic ligand.

The term “oral mucositis” shall also include stomatitis, small intestine-titis, large intestine-titis, proctitis, and similar conditions affecting the mucosal lining of the entire gastrointestinal tract, and related conditions.

The term “R groups” means the group of variable substituents designated as “R” attached to the carbon atoms of the macrocycle, i.e., R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀.

The term “saturated, partially saturated or unsaturated cycle or heterocycle” means a fused ring structure in which 2 carbons of the ring are also part of the fifteen-membered macrocyclic ligand in which the ring can contain no double bonds (in the case of a saturated ring structure) or at least one double bond, which may be conjugated or unconjugated with another double bond. The ring structure can contain 3 to 20 carbon atoms, preferably 5 to 10 carbon atoms, which may be heterocyclic. The cyclic can also contain more than one ring.

In addition, the following abbreviations have the following meanings:

AUC Area under the curve bid Twice daily FGF Fibroblast growth factor g, mg, ml, kg Gram, milligram, milliliter, kilogram Gy Gray ip Intraperitoneal ma Milliamp mM Millimolar mm, cm Millimeter, centimeter SEM Standard error of the mean top Topical

Methods for Treating Oral Mucositis

The present invention involves the administration of a ROS scavenger in the treatment of oral mucositis. A ROS scavenger of the present invention is a superoxide dismutase mimetic, a non-proteinaceous molecule that catalyzes the conversion the of the superoxide radical, O₂, to molecular oxygen and hydrogen peroxide. Such a molecule can, like a native superoxide dismutase enzyme, reduce cell injury resulting from superoxide radical found in diseases involving oxidative stress such as inflammation (Salvemini et al, Arthritis & Rheumatism 44:2909-2921, 2001). The ROS scavengers of the present invention can be pentaaza-macrocyclic complexes, and more specifically, those compositions as disclosed in U.S. Pat. Nos. 5,610,293, 5,637,578, 5,874,421, 5,976,498, 6,084,093, 6,180,620, 6,204,259, 6,214,817, 6,245,758, 6,395,725, and 6,525,041, each of which is incorporated herein by reference in its entirety.

The superoxide dismutase mimetics of the present invention can be represented by the following formula:

wherein

(i) R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀ are independently:

(i^(a)) hydrogen; or

(i^(b)) a moiety independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and aralkyl radicals and radicals attached to the α-carbon or α-amino acids heterocyclyl; or

(i^(c)) a moiety independently selected from the group consisting of —OR₁, —NR₁₁R₁₂, —COR₁₁, —CO₂R₁₁, —CONR₁₁R₁₂, —SR₁₁, —SOR₁₁, —SO₂R₁₁, —SO₂NR₁₁R₁₂, —N(OR₁₁)(R₁₂), —P(O)(OR₁₁)(OR₁₂), —P(O)(OR₁₁)(R₁₂), —OP(O)(OR₁₁)(OR₁₂), and substituents attached to the α-carbon of α-amino acids, wherein R₁₁ and R₁₂ are independently hydrogen or alkyl; and

(ii) optionally, one or more of R₁ or R′₁ and R₂ or R′₂, R₃ or R′₃ and R₄ or R′₄, R₅ or R′₅ and R₆ or R′₆, R₇ or R′₇ and R₈ or R′₈, R₉ or R′₉ and R₁₀ or R′₁₀ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(iii) optionally, one or more of R₁ and R′₁, R₂ and R′₂, R₃ and R′₃, R₄ and R′₄, R₅ and R′₅, R₆ and R′₆, R₇ and R′₇, R₈ and R′₈, R₉ and R′₉, and R₁₀ and R′₁₀, together with the carbon atom to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(iv) optionally, one or more of R₁₀ or R′₁₀ and R₁ or R′₁, R₂ or R′₂ and R₃ or R′₃, R₄ or R′₄ and R₅ or R′₅, R₆ or R′₆ and R₇ or R′₇, or R₈ or R′₈ and R₉ or R′₉ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted nitrogen containing heterocycle having 3 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(v) optionally, one or more of R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀, together with a different one of R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀, which is attached to a different carbon atom in the macrocyclic ligand may be bound to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(vi) combinations of any of (i) through (v) above.

Thus, the pentaaza-macrocyclic ligand compositions useful in the present invention can have any combinations of substituted or unsubstituted R groups, saturated, partially saturated or unsaturated cyclics, heterocyclics, nitrogen containing heterocycles, or straps as defined above.

M can be a transition metal, preferably Mn²⁺, Mn³⁺, Mn⁴⁺, Mn⁶⁺, Mn⁷⁺, Fe²⁺, Fe³⁺, Fe⁴⁺, Fe⁶⁺, Ni²⁺, Ni³⁺, Cu¹⁺, Cu²⁺, V²⁺, V³⁺, V⁴⁺, or V⁵⁺. X, Y and Z can independently be selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof; or

X, Y and Z are independently selected from the group consisting of charge-neutralizing anions which are derived from any monodentate or polydentate coordinating ligand and a ligand system and the corresponding anion thereof; or

X, Y and Z are independently attached to one or more of R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₈, R₇, R′₇, R₈, R′₈, R₉, R′₉, R₁₀, and R′₁₀. n is preferably an integer from 0 to 3.

Alternatively, the superoxide dismutase mimetic can be represented by the formula:

wherein

(i) a nitrogen of the macrocycle and two adjacent carbon atoms to which the nitrogen is attached independently form a substituted or unsubstituted, saturated, partially saturated or unsaturated nitrogen-containing heterocycle W having 2 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(ii) one or more of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀ are independently:

(ii^(a)) hydrogen; or

(ii^(b)) a moiety independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, heterocyclyl, and aralkyl radicals and radicals attached to the α-carbon or α-amino acids; or

(ii^(c)) a moiety independently selected from the group consisting of —OR₁, —NR₁₁R₁₂, —COR₁₁, —CO₂R₁₁, —CONR₁₁R₁₂, —SR₁₁, —SOR₁₁, —SO₂R₁₁, —SO₂NR₁₁R₁₂, —N(OR₁₁)(R₁₂), —P(O)(OR₁₁)(OR₁₂), —P(O)(OR₁₁)(R₁₂), —OP(O)(OR₁₁)(OR₁₂), and substituents attached to the α-carbon of α-amino acids, wherein R₁₁ and R₁₂ are independently hydrogen or alkyl; and

(iii) optionally, one or more of R₁ and R₂ or R′₂, R₃ or R′₃ and R₄ or R′₄, R₅ or R′₅ and R₆ or R′₆, R₇ or R′₇ and Ra or R′₈, R₉ or R′₉ and R₁₀ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(iv) optionally, one or more of R₂ and R′₂, R₃ and R′₃, R₄ and R′₄, R₅ and R′₅, R₆ and R′₆, R₇ and R′₇, R₈ and R′₈, and R₉ and R′₉, together with the carbon atom to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(v) optionally, one or more of R₂ or R′₂ and R₃ or R′₃, R₄ or R′₄ and R₅ or R′₅, R₆ or R′₆ and R₇ or R′₇, or R₈ or R′₈ and R₉ or R′₉ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted nitrogen containing heterocycle having 3 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(vi) optionally, one or more of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀, together with a different one of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀, which is attached to a different carbon atom in the macrocyclic ligand may be bound to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(vii) optionally, one or more of R₁, R₂, R′₂, R₃, R′₃, R₄, R′₄, R₅, R′₅, R₆, R′₆, R₇, R′₇, R₈, R′₈, R₉, R′₉, and R₁₀, may be bound to an atom of heterocycle W to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(viii) combinations of any of (i) through (vii) above.

Thus, the pentaaza-macrocyclic ligand compositions useful in the present invention can have any combinations of substituted or unsubstituted R groups, saturated, partially saturated or unsaturated cyclics, heterocyclics, nitrogen containing heterocycles, or straps as defined above, which may or may not independently connect the W loop and the pentaaza macrocycle.

M can be a transition metal, preferably Mn²⁺, Mn³⁺, Mn⁴⁺, Mn⁶⁺, Mn⁷⁺, Fe²⁺, Fe³⁺, Fe⁴⁺, Fe⁶⁺, Ni²⁺, Ni³⁺, Cu¹⁺, Cu²⁺, V²⁺, V³⁺, V⁴⁺, or V⁵⁺. X, Y and Z can independently be selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof. n is preferably an integer from 0 to 3. W can be a substituted or unsubstituted pyridino moiety.

In another alternative, the superoxide dismutase mimetic can be represented by the formula:

wherein

(i) a nitrogen of the macrocycle and two adjacent carbon atoms to which the nitrogen is attached independently form a substituted or unsubstituted, saturated, partially saturated or unsaturated nitrogen-containing heterocycle W having 2 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(ii) two sets of two adjacent carbon atoms of the macrocycle independently form substituted or unsubstituted, saturated, partially saturated or unsaturated, cycles or heterocycles U and V having 3 to 20 carbon atoms; and

(iii) R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀ are independently:

(iii^(a)) hydrogen; or

(iii^(b)) a moiety independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, heterocyclyl, and aralkyl radicals and radicals attached to the α-carbon or α-amino acids; or

(iii^(c)) a moiety independently selected from the group consisting of —OR₁, —NR₁₁R₁₂, —COR₁₁, —CO₂R₁₁, —CONR₁₁R₁₂, —SR₁₁, —SOR₁₁, —SO₂R₁₁, —SO₂NR₁₁R₁₂, —N(OR₁₁)(R₁₂), —P(O)(OR₁₁)(OR₁₂), —P(O)(OR₁₁)(R₁₂), —OP(O)(OR₁₁)(OR₁₂), and substituents attached to the α-carbon of α-amino acids, wherein R₁, and R₁₂ are independently hydrogen or alkyl; and

(iv) optionally, one or more of R₁ and R₂ or R′₂, R₅ or R′₅ and R₆ or R′₆, R₉ or R′₉ and R₁₀ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(v) optionally, one or more of R₂ and R′₂, R₅ and R′₅, R₆ and R′₆, and R₉ and R′₉, together with the carbon atom to which they are attached independently form a substituted or unsubstituted and saturated, partially saturated, or unsaturated cycle or heterocycle having 3 to 20 carbon atoms; and

(vi) optionally, one or more of R₂ or R′₂ and R₃, R₄ and R₅ or R′₅, R₆ or R′₆ and R₇, or R₈ and R₉ or R′₉ together with the carbon atoms to which they are attached independently form a substituted or unsubstituted nitrogen containing heterocycle having 3 to 20 carbon atoms, which may be an aromatic heterocycle in which case the hydrogen attached to the nitrogen which is both part of the heterocycle and the macrocycle and the R groups attached to the carbon atoms which are both part of the heterocycle and the macrocycle are absent; and

(vii) optionally, one or more of R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀, together with a different one of R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀, which is attached to a different carbon atom in the macrocyclic ligand may be bound to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(viii) optionally, one or more of R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀, may be individually bound to an atom of heterocycles U, V and W to form a strap represented by the formula: —(CH₂)_(I)-Q-(CH₂)_(J)—R—(CH₂)_(K)—S—(CH₂)_(L)—

wherein

I, J, K and L independently are integers from 0 to 10 and Q, R and S are independently selected from the group consisting of alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, aza, amide, ammonium, oxa, thia, sulfonyl, sulfinyl, sulfonamide, phosphoryl, phosphinyl, phosphino, phosphonium, keto, ester, alcohol, carbamate, urea, thiocarbonyl, borates, boranes, boraza, silyl, siloxy, silaza, and combinations thereof; and

(ix) combinations of any of (i) through (viii) above;

Thus, the pentaaza-macrocyclic ligand compositions useful in the present invention can have any combinations of substituted or unsubstituted R groups, saturated, partially saturated or unsaturated cyclics, heterocyclics, nitrogen containing heterocycles, or straps as defined above, which may or may not independently connect the W, U or V loops and the pentaaza macrocycle.

M can be a transition metal, preferably Mn²⁺, Mn³⁺, Mn⁴⁺, Mn⁶⁺, Mn⁷⁺, Fe²⁺, Fe³⁺, Fe⁴⁺, Fe⁶⁺, Ni²⁺, Ni³⁺, Cu¹⁺, Cu²⁺, V²⁺, V³⁺, V⁴⁺, or V⁵⁺. X, Y and Z can independently be selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof. n is preferably an integer from 0 to 3. W can be a substituted or unsubstituted pyridino moiety. U and V can be independently saturated cycloalkyl heterocycles having 3 to 20 carbon atoms, more preferably 4 to 10 carbon atoms, still more preferably trans-cyclohexanyl fused rings. U and V can be trans-cyclohexanyl fused rings while W is a substituted pyridino moiety.

In certain embodiments, the superoxide dismutase mimetic can be the compound identified as M40403, which can be represented by the formula:

The present invention can involve administration of a ROS scavenger, in particular, M40403 or a suitable derivative or analog thereof as described above, to a patient in need thereof. Administration of the ROS scavenger, in particular M40403, can be by numerous routes of administration well known to those of skill in the art.

Administration of the ROS scavenger can be by any suitable route of administration such as, for example, oral, buccal, sublingual, intranasal, inhalation, rectal, intravaginal, transdermal, intradermal, subcutaneous, intramuscular, intraperitoneal, intravenous, intraarterial, intrasternal, intrathecal and the like.

Pharmaceutically acceptable formulations for parenteral or nonparenteral drug delivery are known in the art such as, for example, are set forth in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing (1990). For pharmaceutical compositions and methods of treatment disclosed herein, dosage forms and administration regimes can be determined using standard methods known to skilled artisans, for example as set forth in standard references such as Remington: the Science and Practice of Pharmacy (Alfonso R. Gennaro ed. 19th ed. 1995); Hardman, J. G., et al., Goodman & Gilman's The Pharmacological Basis of Therapeutics, Ninth Edition, McGraw-Hill, 1996; and Rowe, R. C., et al., Handbook of Pharmaceutical Excipients, Fourth Edition, Pharmaceutical Press, 2003.

Pharmaceutical compositions can be formulated to be compatible with the intended route of administration. Solutions or suspensions used for parenteral, intradermal or subcutaneous application can include: a sterile diluent, such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents, such as benzyl alcohol or methyl parabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity, such as sodium chloride or dextrose. Suitable carriers include physiological saline, bacteriostatic water, Cremophor® EL (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). The compositions can be stable during manufacture and storage and preserved against contamination from microorganisms, such as bacteria and fungi.

Proper fluidity can be maintained, for example, by using a coating such as lecithin; by maintaining the required particle size in the case of dispersion, and by using surfactants. Various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal, can control microorganism contamination. Isotonic agents, such as sugars, polyalcohols such as manitol, sorbitol, and sodium chloride can be included in the composition. Compositions that delay absorption can be prepared by including such agents as aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the active compound (e.g., an SCMP) in an appropriate solvent with one or more ingredient, followed by sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and any other required ingredients. Sterile powders for the preparation of sterile injectable solutions include vacuum- and freeze-drying that yield a powder containing the active ingredient and any desired ingredient from a sterile solution. The concentration of active drug, i.e. the ROS scavenger, can be from about 0.1% to about 90% by weight, from about 5% to about 20% by weight, from about 5% to about 17% by weight, from about 8% to about 14% by weight or, in certain embodiments, about 10% by weight.

Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included. Tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, primogel, or corn starch; a lubricant such as magnesium stearate or sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. The concentration of active drug, i.e. the ROS scavenger, can be from about 0.1% to about 99% by weight, from about 5% to about 95% by weight, from about 10% to about 90% by weight, from about 15% to about 85% by weight, from about 20% to about 80% by weight, from about 25% to about 75% by weight, from about 30% to about 70% by weight, from about 35% to about 64% by weight or from about 40% to about 60% by weight.

Administration by inhalation, can be by aerosol spray from a nebulizer or a pressurized container that contains a suitable propellant, e.g., a gas such as carbon dioxide.

Systemic administration can also be transmucosal or transdermal. For transmucosal or transdermal administration, penetrants that can permeate the target barrier(s) are selected. Transmucosal penetrants include detergents, bile salts and fusidic acid derivatives. Nasal sprays or suppositories can be used for transmucosal administration. For transdermal administration, the active compounds are formulated into ointments, salves, gels or creams.

The compounds can also be prepared as suppositories (with bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

In various embodiments, the active compounds can be prepared with carriers that protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid (Alza Corporation; Mountain View, Calif. and Nova Pharmaceuticals, Inc.; Lake Elsinore, Calif.). Liposomal suspensions can also be used as pharmaceutically acceptable carriers (Eppstein, 1985).

Oral formulations or parenteral compositions in unit dosage form can be created to facilitate administration and dosage uniformity. Unit dosage form refers to physically discrete units suited as single doses for a subject to be treated, containing a therapeutically effective quantity of active compound in association with the required pharmaceutical carrier. The specification for unit dosage forms are dictated by, and directly dependent on, the unique characteristics of the active compound and the particular desired therapeutic effect, and the inherent limitations of compounding the active compound.

A typical dose of the ROS scavenger can be from about 0.1 mg up to about 1000 mg or from about 0.001 up to about 10 mg/kg body weight. Doses of 5 mg/kg and 10 mg/kg administered intraperitoneally have shown to produce beneficial effects in rats treated with type II collagen to induced arthritis (Salvemini et al., Arthritis & Rheumatism, 44:2909-2921, 2001). A dose of 2 mg/kg produced less of an effect that was nevertheless significantly different from that in placebo animals. Low doses of a ROS scavenger can be doses of less than about 5 mg/kg or doses equal to or less than about 2 mg/kg body weight, in particular, a dose of about 0.1 mg, about 0.2 mg, about 0.5 mg, about 0.8 mg, about 1 mg, about 2 mg, about 5 mg, about 8 mg, about 10 mg, about 20 mg, about 50 mg, about 80 mg, about 100 mg or about 200 mg or about 0.001 mg/kg, about 0.002 mg/kg, about 0.005 mg/kg, about 0.01 mg/kg, about 0.02 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg, about 0.2 mg/kg, about 0.5 mg/kg, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg or about 4 mg/kg body weight.

Total daily doses of the ROS scavenger can be administered in single or divided doses and in amounts such as, for example, from about 1 to about 2 mg/kg body weight daily and more usually about 0.05 to 1 mg/kg. Dosage unit compositions may contain such amounts of submultiples thereof to make up the total dose. However, one skilled in the art will recognize that the total dosage will vary on the particular composition the particular ROS scavenger administered.

Individuals receiving treatment are, typically, human patients, however, patients receiving treatment can also be animal including companion animal such as dogs and cats, farm animal such as cows, horses, swine as well as birds and exotic animal such as zoo animals.

The amount of active ingredients that may be combined with the carrier materials to produce a single dosage form can vary depending upon the host treated and the particular mode of administration. It will be appreciated that the unit content of active ingredients contained in an individual dose of each dosage form need not in itself constitute an effective amount, as the necessary effective amount could be reached by administration of a number of individual doses. The selection of dosage depends upon the dosage form utilized, the condition being treated, and the particular purpose to be achieved according to the determination of those skilled in the art.

The dosage regimen for treating a disease condition with the compounds and/or compositions of this invention can be selected in accordance with a variety of factors, including the type, age, weight, sex, diet and medical condition of the patient, the route of administration, pharmacological considerations such as the activity, efficacy, pharmacokinetic and toxicology profiles of the particular compound employed, whether a drug delivery system is utilized and whether the compound is administered as part of a drug combination. Thus, the dosage regimen actually employed can, therefore, can deviate from the preferred dosage regimen set forth above.

In various embodiments, the present invention can also involve kits. Such kits can include pharmaceutical compositions and, in addition in certain embodiments, instructions for administration. When supplied as a kit, the different components of the composition can be packaged in separate containers and admixed immediately before use. Such packaging of the components separately can, in certain instances, permit long-term storage without losing activity of the components. In addition, if more than one route of administration is intended or more than one schedule for administration is intended, the different components can be packaged separately and not mixed prior to use. In various embodiments, the different components can be packaged in one composition for administration together.

Kits may also include reagents in separate containers such as, for example sterile water or saline to be added to a lyophilized active component packaged separately. For example, sealed glass ampules may contain lyophilized ROS scavenger, sterile water, sterile saline or sterile each of which has been packaged under a neutral non-reacting gas, such as nitrogen. Ampules may consist of any suitable material, such as glass, organic polymers, such as polycarbonate, polystyrene, etc., ceramic, metal or any other material typically employed to hold reagents. Other examples of suitable containers include bottles that may be fabricated from similar substances as ampules, and envelopes that may consist of foil-lined interiors, such as aluminum or an alloy. Other containers include test tubes, vials, flasks, bottles, syringes, etc. Containers may have a sterile access port, such as a bottle having a stopper that can be pierced by a hypodermic injection needle. Other containers may have two compartments that are separated by a readily removable membrane that upon removal permits the components to mix. Removable membranes may be glass, plastic, rubber, etc.

In certain embodiments, kits can be supplied with instructional materials. Instructions may be printed on paper or other substrate, and/or may be supplied as an electronic-readable medium, such as a floppy disc, mini-CD-ROM, CD-ROM, DVD-ROM, Zip disc, videotape, audio tape, etc. Detailed instructions may not be physically associated with the kit; instead, a user may be directed to an internet web site specified by the manufacturer or distributor of the kit, or supplied as electronic mail.

EXAMPLES

Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following specific examples are offered by way of illustration and not by way of limiting the remaining disclosure.

Example 1

Objective

The objective of this study is to evaluate the effect of M40403, administered by either ip or topical routes, with 2 different schedules, on the frequency, severity and duration of oral mucositis induced in hamsters by acute radiation.

Summary

M40403 was given as an intraperitoneal (ip) injection at a 30 mg/kg/dose twice daily (a) from day −1 to day 15, or (b) day −1 to day 3. M40403 was also given by ip injection at a dose of 3 mg/kg twice daily from day −1 to day 15. In addition, M40403 was given as a topical dose directed to the buccal mucosa in 0.2 ml doses of either 3 mg/ml or 30 mg/ml twice daily from day −1 to day 15. The greatest reduction in oral mucositis was seen in the group treated with M40403 at 30 mg/kg/dose ip twice daily from day −1 to day 3. This group had a statistically significant reduction in the number of animal days with a mucositis score of 3 or higher (P<0.001) and significantly lower mean mucositis scores than the control group on days 18 (P=0.041), 20 (P<0.001), 22 (P<0.001), 24 (P<0.001) and 26 (P=0.002). No other treatment groups in this study showed a reduction in mucositis either by long term (day-1 to day 15) ip dosing or by topical dosing. This study establishes a schedule-dependent dosing method for the treatment of oral mucositis with M40403.

Acute Radiation Model

The acute radiation model in hamsters, developed by the Principal Investigator, has proven to be an accurate, efficient and cost-effective technique to provide a preliminary evaluation of anti-mucositis compounds (9). The course of mucositis in this model is well defined and results in peak scores approximately 14-16 Days following radiation. The acute model has little systemic toxicity, resulting in few hamster deaths, thus permitting the use of smaller groups (N=7-8) for initial efficacy studies. It has also been used to study specific mechanistic elements in the pathogenesis of mucositis. Molecules that show efficacy in the acute radiation model may be further evaluated in the more complex models of fractionated radiation, chemotherapy, or concomitant therapy.

In this study, an acute radiation dose of 40 Gy on day 0 was administered locally to the cheek pouch. Clinically significant mucositis was observed on days 12 through 28.

Protocol Summary

Forty-eight male Syrian Golden Hamsters were divided randomly into 6 groups of 8 animals and given an acute radiation dose of 40 Gy directed to their left buccal cheek pouch. This was accomplished by anesthetizing the animals and everting the left buccal pouch, while protecting the rest of the animal with a lead shield. Test materials were given topically or by ip injection twice daily as detailed in Table 1. Mucositis was evaluated clinically starting on Day 6, and continued on alternate days until day 28.

Evaluation

Mucositis Evaluation

The grade of mucositis was scored, beginning on day 6, and for every second day thereafter, through and including day 28 (for scoring scheme, see Table 2 and FIG. 2 ). The effect on mucositis of each drug treatment compared to ip vehicle-treated controls was assessed according to the following parameters:

The difference in the number of days hamsters in each group have ulcerative (score ≥3) mucositis.

On each evaluation day, the number of animals with a blinded mucositis score of ≥3 in each drug treatment group was compared to the vehicle-treated control group. Differences were compared on a cumulative basis and statistical significance was determined by chi-square analysis. Efficacy, in this analysis, is defined as a significant reduction in the number of days that a group of animals had ulcerations (scores ≥3) when compared to the control group.

TABLE 2 Table 2 ACT-02: Mucositis Scoring. Score: Description: 0 Pouch completely healthy. No erythema or vasodilation. 1 Light to severe erythema and vasodilation. No erosion of mucosa. 2 Severe erythema and vasodilation. Erosion of superficial aspects of mucosa leaving denuded areas. Decreased stippling of mucosa. 3 Formation of off-white ulcers in one or more places. Ulcers may have a yellow/gray due to pseudomembrane. Cumulative size of ulcers should equal about ¼ of the pouch. Severe erythema and vasodilation. 4 Cumulative seize of ulcers should equal about ½ of the pouch. Loss of pliability. Severe erythema and vasodilation. 5 Virtually all of pouch is ulcerated. Loss of pliability (pouch can only partially be extracted from mouth).

Rank Sum Differences in Daily Mucositis Scores.

For each evaluation day the scores of the control group were compared to those of the treated groups using non-parametric rank sum analysis. Treatment success was considered as a statistically significant lowering of scores in the treated group on 2 or more days from day 6 to day 28.

Weights and Survival

All animals were weighed daily and their survival recorded, in order to assess possible differences in animal weight among treatment groups as an indication for mucositis severity and/or possible toxicity resulting from the treatments.

Study Design

All forty-eight (48) male Syrian Golden Hamsters were given an acute radiation dose of 40 Gy directed to their left buccal cheek pouch. This was accomplished by anesthetizing the animals and everting the left buccal pouch, while protecting the rest of the animal with a lead shield. Test materials were given either topically or by ip injection twice daily as detailed in Table 1. Mucositis was evaluated clinically starting on day 6, and continuing on alternate days until day 28. The study timeline is depicted in FIG. 1 . Dose levels were determined based on the results of a previous acute toxicity study (data not shown), and the levels shown in Table 1.

TABLE 1 Table 1. ACT-02. Study Design Group Number of Treatment Volume Number Animals Treatment Schedule* (mL) 1 8 males Vehicle, ip, bid Day −1 to 15 Adjust per body weigh 2 8 males M40403, ip, bid Day −1 to 15 Adjust per 3 mg/kg body weigh 3 8 males M40403, ip, bid Day −1 to 15 Adjust per 30 mg/kg body weigh 4 8 males M40403, topical, bid Day −1 to 15 0.2 ml per 3 mg/ml dose 5 8 males M40403, topical, bid Day −1 to 15 0.2 ml per 30 mg/ml dose 6 8 males M40403, ip, bid Day −1 to 3 Adjust per 30 mg/kg body weigh *The first close on day 0 was performed 30 minutes prior to radiation. The second dose was given at least 4 hours after radiation.

At the end of the study on day 28, the animals in group 1 were used in a modified pK study as follows: Two animals were injected with vehicle and blood was drawn 30 minutes after the injection. Six animals were injected with 30 mg/kg of M40403. At 15, 30 and 90 minutes post injection, 2 animals from this group were sacrificed and blood was obtained by cardiac puncture. Blood was collected in lithium heparin, kept on ice for 30 minutes and the plasma was obtained after centrifugation. Plasma was transferred to labeled tubes, snap frozen in liquid nitrogen and shipped to ActivBiotics.

Material and Methods

Location of Study Performance

The study was performed at Biomodels AAALAC accredited facility in Cambridge Mass. The IACUC approval number 04-0624-2 for this study was obtained from Biomodels IACUC.

Animals

Male LVG Syrian Golden Hamsters (Charles River Laboratories), aged 5 to 6 weeks, with average body weight of 84.1 g at study commencement, were used. Animals were individually numbered using an ear punch and housed in small groups of approximately 8 animals per cage. Animals were acclimatized prior to study commencement. During this period of 2 days, the animals were observed daily in order to reject animals that presented in poor condition.

Housing

The study was performed in animal rooms provided with filtered air at a temperature of 70° F.+/−5° F. and 50%+/−20% relative humidity. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour. The room was on an automatic timer for a light/dark cycle of 12 hours on and 12 hours off with no twilight. Bed-O-Cobs® bedding was used. Bedding was changed a minimum of once per week. Cages, tops, bottles, etc. were washed with a commercial detergent and allowed to air dry. A commercial disinfectant was used to disinfect surfaces and materials introduced into the hood. Floors were swept daily and mopped a minimum of twice weekly with a commercial detergent. Walls and cage racks were sponged a minimum of once per month with a dilute bleach solution. A cage card or label with the appropriate information necessary to identify the study, dose, animal number and treatment group marked all cages. The temperature and relative humidity was recorded during the study, and the records were retained.

Diet

Animals were fed with a Purina Labdiet® 5061 rodent diet and water was provided ad libitum. Animal randomization and allocations.

Animals were randomly and prospectively divided into eight (8) treatment groups prior to irradiation. Each animal was identified by an ear punch corresponding to an individual number. For more consistent identification, ear punch numbering was used rather than tagging, since tags may become dislodged during the course of the study. A cage card was used to identify each cage and was marked with the study number, treatment group number and animal numbers.

Dosing and Drug Application

Dosing solutions were made immediately prior to use. Aseptic technique was used for all preparation procedures. A 26-mM sodium bicarbonate buffer solution was prepared as the vehicle. The resultant pH was approximately 8.1 to 8.3.

Mucositis Induction

Mucositis was induced using a standardized acute radiation protocol. A single dose of radiation (40 Gy/dose) was administered to all animals on day 0. Radiation was generated with a 250 kilovolt potential (15-ma) source at a focal distance of 50 cm, hardened with a 0.35 mm Cu filtration system. Irradiation targeted the left buccal pouch mucosa at a rate of 3.2 Gy/minute. Prior to irradiation, animals were anesthetized with an intra-peritoneal injection of Ketamine (160 mg/kg) and Xylazine (8 mg/kg). The left buccal pouch was everted, fixed and isolated using a lead shield.

Mucositis Scoring

The mucositis score, weight change and survival were measured throughout the study as described above. For the evaluation of mucositis, the animals were anesthetized with isoflurane and the left pouch everted. Mucositis was scored visually by comparison to a validated photographic scale (FIG. 2 ), ranging from 0 for normal, to 5 for severe ulceration (clinical scoring). In descriptive terms, this scale is defined as follows:

A score of 1-2 is considered to represent a mild stage of the disease, whereas a score of 3-5 is considered to indicate moderate to severe mucositis. Following visual scoring, a photograph was taken of each animal's mucosa using a standardized technique. At the conclusion of the experiment, all films were developed and the photographs randomly numbered. At least two independent trained observers graded the photographs in blinded fashion using the above described scale (blinded scoring).

Results and Discussion

Survival

Five deaths occurred during this study. Two animals, hamsters 8 and 23, in the vehicle control group and the group treated with M40403 at 30 mg/kg twice a day from day −1 to day 15 died on day 0 from apparent anesthesia overdoses. In addition, two animals (hamsters 45 and 46) died in the group treated with M40403 at 30 mg/kg twice daily from day −1 to day 3. Hamster 45 died during radiation and hamster 46 was found dead the following morning. Both deaths appeared to be the result of anesthesia overdose, in the case of hamster 46 the response was delayed. The occurrence of anesthesia deaths in this model is anticipated in the experimental design. Hamster 19, in the group treated ip with M40403 at 30 mg/kg twice a day from day −1 to day 15 died on day 12. This death resulted after an extended period of failure to gain weight and may reflect toxicity of this dosing regimen as all animals in this group showed lack of weight gain (see FIGS. 3 and 4 below).

Weight Change (FIGS. 3 and 4 ).

The percent daily weight change for each group is shown in FIG. 3 . The groups treated by ip injection were groups 1, 2, 3 and 6. The control animals (group 1) gained an average of 64.0% of their starting weights by the end of the study. The animals treated ip with M40403 at 3 mg/kg/dose twice daily from day −1 to day 15 (group 2) gained an average of 66.0% of their starting weight during the study. The group treated ip with M40403 at 30 mg/kg/dose twice daily from day −1 to day 15 (group 3) gained an average of 33.9% of their starting weight by day 28. The group treated ip with M40403 at 30 mg/kg/dose twice daily from day 1 to day 3 gained an average of 54.7% of their starting weight during the study. In both groups treated by ip injection with 30 mg/kg M40403, there was a general lack of weight gain during dosing.

In the 2 groups treated topically with M40403 (groups 4 and 5) there was no apparent change in weight gain when compared to the control group. The animals treated topically with M40403 at 3 mg/ml twice daily from day −1 to day 15 gained an average of 65.8% of their starting weight by day 28. The group treated topically with M40403 at 30 mg/ml twice daily from day −1 to day 15 gained an average of 71.9% of their starting weight during the study.

The significance of these differences was evaluated by calculating the mean area under the curve for the percentage weight gain for each animal and comparing the groups using a OneWay ANOVA test. There was a significant difference between the control group and group 3 treated by ip injection with M40403 twice daily (30 mg/kg) on days −1 to 15 (P<0.001). There was also a significant difference between the control group and the group treated by ip injection with M40403 twice daily at 30 mg/kg/dose from day −1 to day 3 (P=0.018). No other significant differences were seen. The topical treatment groups (groups 4 and 5) showed no significant weight differences when compared with the control group. The AUC data is shown in FIG. 4 . Many of the groups with a significant reduction in weight gain are the same groups in which animal deaths were observed.

TABLE 4 Day Group Comparison 6 8 10 12 14 16 18 20 22 24 26 28 Control v 0.099 0.630 0.950 0.286 0.983 0.630 0.436 0.571 0.754 0.325 0.516 0.070 M40403 ip 3 mg/kg day −1 to 15 Control v 0.055 0.981 0.981 0.055 0.856 0.938 0.221 0.622 0.698 0.067 0.979 0.225 M40403 ip 30 mg/kg day −1 to 15 Control v 0.099 0.099 0.630 0.463 0.463 0.438 0.983 0.983 0.163 0.632 0.517 0.139 M40403 top 3 mg/ml day −1 to 15 Control v 0.630 0.099 0.002 0.573 0.465 0.571 0.884 0.884 0.573 0.632 0.754 0.884 M40403 top 30 mg/ml day −1 to 15 Control v 0.127 0.059 0.546 0.936 0.221 0.312 0.041 <0.001 <0.001 <0.001 0.002 0.551 M40403 ip 30 mg/kg day −1 to 3 Table 4. ACT-02. The significance of group differences observed in daily mucositis scores was determined using the Mann-Whitney rank sum test. This nonparametric statistic is appropriate for the visual mucositis scoring scale. The p values for each calculation are shown.

Mucositis (FIGS. 5 & 6 , Tables 3 & 4)

Mean daily mucositis scores for each group are shown in FIG. 5 . The groups treated by ip injection were groups 1, 2, 3 and 6. The control animals (group 1) had a peak mean mucositis score of 2.8 which occurred on day 16. The animals treated with M40403 at 3 mg/kg/dose ip twice daily from day −1 to day 15 (group 2) had a peak mucositis score of 2.8 on day 18, and the overall progression of mucositis severity in this group was very similar to that observed in the control group. The group treated with M40403 at 30 mg/kg/dose twice daily from day −1 to day 15 (group 3) had a peak mucositis score of 3.0 on day 18 and had a mucositis progression that was indistinguishable from that observed in groups 1 and 2. The group treated with M40403 at 30 mg/kg/dose ip twice daily from day −1 to day 3 had a peak mucositis score of 3.0 on day 14, but from day 16 to day 28, the mucositis severity in this group decreased much more rapidly than the control group suggesting that this schedule of treatment was efficacious.

In the 2 groups treated topically with M40403 (groups 4 and 5) there was no apparent effect of treatment on the course of mucositis. The animals treated topically with M40403 at 3 mg/ml twice daily from day −1 to day 15 had mucositis scores that closely paralleled those in the control group. The group treated topically with M40403 at 30 mg/ml twice daily from day −1 to day 15 showed a reduction of mucositis severity on day 10. However, from day 12 to the end of the study, the scores in this group were only slightly less than those in the control group.

The significance of the reductions in the mucositis scores seen in the groups treated with M40403 were evaluated by calculating the percentage of animal days with a score of 3 or higher. The results of this analysis are shown in FIG. 6 and Table 3. In the control group, the percentage of animal days with a mucositis score of 3 or higher was 31%. Treatment by ip injection with M40403 at 3 mg/kg/dose twice daily from day −1 to day 15 resulted in a percentage of animal days with a score of 3 or higher of 34%. Treatment by ip injection with M40403 at 30 mg/kg/dose twice daily from day to −1 to day 15 resulted in 37% percent of animal days with a score of 3 or higher. Treatment by ip injection with M40403 at 30 mg/kg twice daily from day −1 to day 3 (group 6) resulted in a decrease in the number of animal days with a score of 3 or higher to 17%.

TABLE 3 Chi Sq Group Days >= 3 Days < 3 Total Days % Days >= 3 v control P Value Vehicle ip control 52 116 168 0.31 — — day −1 to 15 M40403 ip 3 mg/kg 66 126 192 0.34 0.3440 0.563 day −1 to 15 M40403 ip 30 mg/kg 56 96 152 0.37 0.9890 0.320 day −1 to 15 M40403 top 3 mg/ml 73 119 192 0.38 1.6760 0.196 day −1 to 15 M40403 top 30 mg/ml 64 128 192 0.33 0.136 0.712 day −1 to 15 M40403 ip 30 mg/kg 24 120 144 0.17 7.83 0.005 day −1 to 3 Table 3. ACT-02. Chi-square analysis of the total number of days the animals in each group spent with a mucositis score of 3 or more. This statistic is a measure of severity of ulceration, a clinically important outcome.

In the animals receiving M40403 topically, there were no apparent reductions of mucositis severity at either dosing level. Topical treatment with M40403 at 3 mg/ml twice daily from day −1 to day 15 resulted in an increase in the number of animal days with a score of 3 or higher to 38%. Topical treatment with M40403 at 30 mg/ml twice daily from days −1 to 15 resulted in 33% of animal days with a score of 3 or higher.

When compared using a chi-squared test, the group treated by ip injection with M40403 at 30 mg/kg/dose twice daily on days −1 to day 3 had the greatest reduction in mucositis and significantly fewer days with a score of 3 or higher (P=0.005). No other study group exhibited a reduction in mucositis severity and no group demonstrated a significant difference in mucositis severity by this analysis.

Further analysis of the mucositis scores was performed by comparing the scores for the M40403 treated groups with the control on each day of scoring using the Mann-Whitney Rank Sum test. The results of this analysis are shown in Table 4. The group treated by ip injection with M40403 at 30 mg/kg/dose twice daily on days −1 to day 3 (group 6) had the greatest reduction in mucositis and significantly lower scores than the control group on days 18 (P=0.041), 20 (P<0.001), 22 (P<0.001), 24 (P<0.001) and 26 (P=0.002). No other study group had more than a single day showing a significant lowering of mucositis severity. Given that group 2 received the same dose of drug for a longer duration, these results suggest that the schedule of dosing M40403 is crucial in establishing an effective treatment for mucositis.

Conclusions

1. Based on observations of mortality and body weight, ip injections of M40403, twice daily, at 30 mg/kg appear to show some toxicity. The longer dosing in group 3 resulted in mortality and significant weight loss. When dosed for the shorter period of day −1 to day 3 (group 6), the weight loss was reversed after the cessation of dosing.

2. Topical dosing of M40403 did not appear to have any effect on either weight or mucositis severity. It appears the topical dosing, in the formulation used here, is an ineffective method of delivery of M40403 in this model.

3. The group treated by ip injection with M40403 at 30 mg/kg/dose twice daily on days 1 to 3 had a statistically significant reduction in the number of animal days with a mucositis score of 3 or higher (P=0.005) and significantly lower mucositis scores than the control group on days 18 (P=0.041), 20 (P<0.001), 22 (P<0.001), 24 (P<0.001) and 26 (P=0.002). This result suggests that M40403 may be an effective agent in the treatment of oral mucositis. Further studies of dose and schedule may be required to optimize the efficacy of M40403.

Example 2—Repeat of Day −1 to 3 of 30 mg/kg IV of Example 1 Study

Acute Radiation Model

The acute radiation model in hamsters, developed by the Principal Investigator, has proven to be an accurate, efficient and cost-effective technique to provide a preliminary evaluation of anti-mucositis compounds (9). The course of mucositis in this model is well defined and results in peak scores approximately 14-16 Days following radiation. The acute model has little systemic toxicity, resulting in few hamster deaths, thus permitting the use of smaller groups (N=7-8) for initial efficacy studies. It has also been used to study specific mechanistic elements in the pathogenesis of mucositis. Molecules that show efficacy in the acute radiation model may be further evaluated in the more complex models of fractionated radiation, chemotherapy, or concomitant therapy.

In this study, an acute radiation dose of 40 Gy on day 0 was administered locally to the cheek pouch. Clinically significant mucositis was observed on days 12 through 28.

Protocol Summary

Seventy-two male Syrian Golden Hamsters were divided randomly into 9 groups of 8 animals and given an acute radiation dose of 40 Gy directed to their left buccal cheek pouch. This was accomplished by anesthetizing the animals and everting the left buccal pouch, while protecting the rest of the animal with a lead shield. Test materials were given topically or by ip injection twice daily as detailed in Table 5. Mucositis was evaluated clinically starting on Day 6, and continued on alternate days until day 28.

TABLE 5 Table 1. ACT-03. Study Design Group Number of Treatment Volume Number Animals Treatment Schedule* (mL) 1 8 males Vehicle, ip, bid Day −1 to 3 Adjust per body weigh 2 8 males M40403, ip, QD Day −1 to 3 Adjust per 30 mg/kg body weigh 3 8 males M40403, ip, bid Day −1 to 3 Adjust per 3 mg/kg body weigh 4 8 males M40403, ip, bid Day −1 to 3 Adjust per 10 mg/kg body weigh 5 8 males M40403, ip, bid Day −1 to 3 Adjust per 30 mg/kg body weigh 6 8 males M40403, ip, QD Day 0 to 3 Adjust per 30 mg/kg body weigh 7 8 males M40403, ip, bid Day 0 to 3 Adjust per 30 mg/kg body weigh 8 8 males M40403, ip, bid Day 0 Adjust per 30 mg/kg body weigh 9 8 males M40403, ip, bid Day 0 and Day 7 Adjust per 30 mg/kg body weigh *For BID dosing the first dose on day 0 will be performed 30 minutes prior to radiation. The second dose will be given at least 4 hours after radiation. For QD dosing, the dose will be administered on day 0 at 30 minutes prior to irradiation.

Evaluation

Mucositis Evaluation

The grade of mucositis was scored, beginning on day 6, and for every second day thereafter, through and including day 28 (for scoring scheme, see Table 6 and FIG. 8 ). The effect on mucositis of each drug treatment compared to ip vehicle-treated controls was assessed according to the following parameters:

The difference in the number of days hamsters in each group have ulcerative (score ≥3) mucositis.

On each evaluation day, the number of animals with a blinded mucositis score of ≥3 in each drug treatment group was compared to the vehicle-treated control group.

Differences were compared on a cumulative basis and statistical significance was determined by chi-square analysis. Efficacy, in this analysis, is defined as a significant reduction in the number of days that a group of animals had ulcerations (scores ≥3) when compared to the control group.

TABLE 6 Table 2 ACT-03: Mucositis Scoring. Score: Description: 0 Pouch completely healthy. No erythema or vasodilation. 1 Light to severe erythema and vasodilation. No erosion of mucosa. 2 Severe erythema and vasodilation. Erosion of superficial aspects of mucosa leaving denuded areas. Decreased stippling of mucosa. 3 Formation of off-white ulcers in one or more places. Ulcers may have a yellow/gray due to pseudomembrane. Cumulative size of ulcers should equal about ¼ of the pouch. Severe erythema and vasodilation. 4 Cumulative seize of ulcers should equal about ½ of the pouch. Loss of pliability. Severe erythema and vasodilation. 5 Virtually all of pouch is ulcerated. Loss of pliability (pouch can only partially be extracted from mouth).

Rank Sum Differences in Daily Mucositis Scores.

For each evaluation day the scores of the control group were compared to those of the treated groups using non-parametric rank sum analysis. Treatment success was considered as a statistically significant lowering of scores in the treated group on 2 or more days from day 6 to day 28.

Weights and Survival

All animals were weighed daily and their survival recorded, in order to assess possible differences in animal weight among treatment groups as an indication for mucositis severity and/or possible toxicity resulting from the treatments.

Study Design

All seventy-two (72) male Syrian Golden Hamsters were given an acute radiation dose of 40 Gy directed to their left buccal cheek pouch. This was accomplished by anesthetizing the animals and everting the left buccal pouch, while protecting the rest of the animal with a lead shield. Test materials were given either topically or by ip injection twice daily as detailed in Table 5. Mucositis was evaluated clinically starting on day 6, and continuing on alternate days until day 28. The study timeline is depicted in FIG. 7 . Dose levels were determined based on the results of a previous acute toxicity study (data not published), and in the prior mucositis study of Example 1, shown in Table 5.

Material and Methods

Location of Study Performance

The study was performed at Biomodels AAALAC accredited facility in Cambridge Mass. The IACUC approval number 04-0624-2 for this study was obtained from Biomodels IACUC.

Animals

Male LVG Syrian Golden Hamsters (Charles River Laboratories), aged 5 to 6 weeks, with average body weight of 92.7 at study commencement, were used. Animals were individually numbered using an ear punch and housed in small groups of approximately 8 animals per cage. Animals were acclimatized prior to study commencement. During this period of 3 days, the animals were observed daily in order to reject animals that presented in poor condition.

Housing

The study was performed in animal rooms provided with filtered air at a temperature of 70° F.+/−5° F. and 50%+/−20% relative humidity. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour. The room was on an automatic timer for a light/dark cycle of 12 hours on and 12 hours off with no twilight. Bed-O-Cobs® bedding was used. Bedding was changed a minimum of once per week. Cages, tops, bottles, etc. were washed with a commercial detergent and allowed to air dry. A commercial disinfectant was used to disinfect surfaces and materials introduced into the hood. Floors were swept daily and mopped a minimum of twice weekly with a commercial detergent. Walls and cage racks were sponged a minimum of once per month with a dilute bleach solution. A cage card or label with the appropriate information necessary to identify the study, dose, animal number and treatment group marked all cages. The temperature and relative humidity was recorded during the study, and the records were retained.

Diet

Animals were fed with a Purina Labdiet® 5061 rodent diet and water was provided ad libitum. Animal randomization and allocations.

Animals were randomly and prospectively divided into eight (8) treatment groups prior to irradiation. Each animal was identified by an ear punch corresponding to an individual number. For more consistent identification, ear punch numbering was used rather than tagging, since tags may become dislodged during the course of the study. A cage card was used to identify each cage and was marked with the study number (ACT-03), treatment group number and animal numbers.

Dosing and Drug Application

Dosing solutions were made immediately prior to use. Aseptic technique was used for all preparation procedures. A 26-mM sodium bicarbonate buffer solution was prepared as the vehicle. The resultant pH was approximately 8.1 to 8.3.

Mucositis Induction

Mucositis was induced using a standardized acute radiation protocol. A single dose of radiation (40 Gy/dose) was administered to all animals on day 0. Radiation was generated with a 250 kilovolt potential (15-ma) source at a focal distance of 50 cm, hardened with a 0.35 mm Cu filtration system. Irradiation targeted the left buccal pouch mucosa at a rate of 3.2 Gy/minute. Prior to irradiation, animals were anesthetized with an intra-peritoneal injection of Ketamine (160 mg/kg) and Xylazine (8 mg/kg). The left buccal pouch was everted, fixed and isolated using a lead shield.

Mucositis Scoring

The mucositis score, weight change and survival were measured throughout the study as described above. For the evaluation of mucositis, the animals were anesthetized with isoflurane and the left pouch everted. Mucositis was scored visually by comparison to a validated photographic scale (FIG. 8 ), ranging from 0 for normal, to 5 for severe ulceration (clinical scoring). In descriptive terms, this scale is defined as follows:

A score of 1-2 is considered to represent a mild stage of the disease, whereas a score of 3-5 is considered to indicate moderate to severe mucositis. Following visual scoring, a photograph was taken of each animal's mucosa using a standardized technique. At the conclusion of the experiment, all films were developed and the photographs randomly numbered. At least two independent trained observers graded the photographs in blinded fashion using the above-described scale (blinded scoring).

Results and Discussion

Survival

Seven deaths occurred during this study, all on the day of radiation (Day 0). Two animals each died in the group treated with M40403 at 30 mg/kg once daily from day −1 to day 3, the group treated with M40403 at 30 mg/kg twice daily from day −1 to day 3 and the group treated with M40403 at 30 mg/kg once daily from day 0 to day 3. One animal died in the group treated with M40403 at 30 mg/kg once daily on day 0 and day 7

Weight Change (FIGS. 9 and 10 ).

The percent daily weight change for each group is shown in FIG. 9 . The control animals (group 1) gained an average of 62.5% of their starting weights by the end of the study. The animals treated ip with M40403 at 30 mg/kg/dose once daily from day −1 to day 3 (group 2) gained an average of 60.7% of their starting weight during the study. The group treated ip with M40403 at 3 mg/kg/dose twice daily from day −1 to day 3 (group 3) gained an average of 59.3% of their starting weight by day 28. The group treated ip with M40403 at 10 mg/kg/dose twice daily from day −1 to day 3 (group 4) gained an average of 56.2% of their starting weight during the study. The group treated ip with M40403 at 30 mg/kg/dose twice daily from day −1 to day 3 (group 5) gained an average of 52.9% of their starting weight during the study. The group treated ip with M40403 at 30 mg/kg/dose once daily from day 0 to day 3 (group 6) gained an average of 54.8% of their starting weight during the study. The group treated ip with M40403 at 30 mg/kg/dose twice daily from day 0 to day 3 (group 7) gained an average of 58.1% of their starting weight during the study. The group treated ip with M40403 at 30 mg/kg/dose twice daily on day 0 (group 8) gained an average of 59.3% of their starting weight during the study. The group treated ip with M40403 at 30 mg/kg/dose twice daily on day 0 and on day 7 (group 9) gained an average of 57.3% of their starting weight during the study.

The significance of these differences was evaluated by calculating the mean area under the curve for the percentage weight gain for each animal and comparing the groups using a One-Way ANOVA test. There were significant differences between the vehicle control group and the group treated with M40403 at 30 mg/kg/dose once daily from day −1 to day 3 (P=0.039), the group treated with M40403 at 3 mg/kg/dose twice daily from day −1 to day 3 (P=0.040), the group treated with M40403 at 10 mg/kg/dose twice daily from day −1 to day 3 (0.018), the group treated with M40403 at 30 mg/kg/dose twice daily from day −1 to day 3 (P<0.001), the group treated with M40403 at 30 mg/kg/dose once daily from day 0 to day 3 (P=0.009) and the group treated with M40403 at 30 mg/kg/dose twice daily from day 0 to day 3 (P=0.002). The results of this analysis are shown in FIG. 10 .

Mucositis (FIGS. 11 & 12 , Tables 7 & 8)

Mean daily mucositis scores for each group are shown in FIG. 11 . The control animals (group 1) had a peak mean mucositis score of 3.1 on day 16. The animals treated ip with M40403 at 30 mg/kg/dose once daily from day −1 to day 3 (group 2) had a peak mean mucositis score of 2.8 on day 16. The group treated ip with M40403 at 3 mg/kg/dose twice daily from day −1 to day 3 (group 3) had a peak mean mucositis score of 2.9, which occurred on days 16 and 18. The group treated ip with M40403 at 10 mg/kg/dose twice daily from day −1 to day 3 (group 4) had a peak mean mucositis score of 3.0 on day 16. The group treated ip with M40403 at 30 mg/kg/dose twice daily from day −1 to day 3 (group 5) had a peak mean mucositis score of 2.8 on day 16. The group treated ip with M40403 at 30 mg/kg/dose once daily from day 0 to day 3 (group 6) had a peak mean mucositis score of 3.2 on day 16. The group treated ip with M40403 at 30 mg/kg/dose twice daily from day 0 to day 3 (group 7) had a peak mean mucositis score of 2.2 on day 16. The group treated ip with M40403 at 30 mg/kg/dose twice daily on day 0 (group 8) had a peak mean mucositis score of 2.4 on day 14. The group treated ip with M40403 at 30 mg/kg/dose twice daily on day 0 and on day 7 (group 9) had a peak mean mucositis score of 2.4 which occurred on days 16 and 18.

The significance of the reductions in the mucositis scores seen in the groups treated with M40403 were evaluated by calculating the percentage of animal days with a score of 3 or higher. The results of this analysis are shown in FIG. 12 and Table 7. In the control group, the percentage of animal days with a mucositis score of 3 or higher was 36.5%. Treatment by ip injection with M40403 at 30 mg/kg/dose once daily from day −1 to day 3 (group 2) or M40403 at 3 mg/kg/dose twice daily from day −1 to day 3 (group 3) resulted in a percentage of animal days with a score of 3 or higher of 25%. Treatment with M40403 at 10 mg/kg/dose twice daily from day −1 to day 3 (group 4) resulted in a percentage of animal days with a score of 3 or higher of 29.2%. Treatment with M40403 at 30 mg/kg/dose twice daily from day −1 to day 3 (group 5) resulted in a percentage of animal days with a score of 3 or higher of 22.2%. Treatment with M40403 at 30 mg/kg/dose once daily from day 0 to day 3 (group 6) resulted in a percentage of animal days with a score of 3 or higher of 30.6%. Treatment with M40403 at 30 mg/kg/dose twice daily from day 0 to day 3 (group 7) resulted in a percentage of animal days with a score of 3 or higher of 9.4%. Treatment with M40403 at 30 mg/kg/dose twice daily on day 0 (group 8) resulted in a percentage of animal days with a score of 3 or higher of 13.5%. Treatment with M40403 at 30 mg/kg/dose twice daily on day 0 and on day 7 (group 9) resulted in a percentage of animal days with a score of 3 or higher of 15.5%. When compared using a chi-squared test, significant reductions in the number of animal days with a score of 3 or higher were seen in the groups treated with M40403 at 30 mg/kg/dose once daily from day −1 to day 3 (group 2, P=0.034), M40403 at 3 mg/kg/dose twice daily from day −1 to day 3 (group 3, P=0.020), M40403 at 30 mg/kg/dose twice daily from day −1 to day 3 (group 5, P=0.007), M40403 at 30 mg/kg/dose twice daily from day 0 to day 3 (group 7, P<0.001), M40403 at 30 mg/kg/dose twice daily on day 0 (group 8, P<0.001) and M40403 at 30 mg/kg/dose twice daily on day 0 and on day 7 (group 9, P<0.001). This data is shown in FIG. 12 and Table 7.

TABLE 7 Chi Sq Group Days >= 3 Days < 3 Total Days % Days >= 3 v control P Value Vehicle ip control bid 70 122 192 36.46% — — day −1 to 3 M40403 ip 30 mg/kg qd 36 108 144 25.00% 4.4860 0.034 day −1 to 3 M40403 ip 3 mg/kg bid 48 144 192 25.00% 5.3950 0.020 day −1 to 3 M40403 ip 10 mg/kg bid 56 136 192 29.17% 1.1996 0.158 day −1 to 3 M40403 ip 30 mg/kg bid 32 112 144 22.22% 7.229 0.007 day −1 to 3 M40403 ip 30 mg/kg qd 44 100 144 30.56% 1.029 0.310 day 0 to 3 M40403 ip 30 mg/kg bid 18 174 192 9.38% 33.3440 <0.001 day 0 to 3 M40403 ip 30 mg/kg bid 26 166 192 13.54% 25.681 <0.001 day 0 M40403 ip 30 mg/kg bid 26 142 168 15.48% 19.113 <0.001 day 0 & 7 Table 3. ACT-03. Chi-square analysis of the total number of days the animals in each group spent with a mucositis score of three or more. This statistic is a measure of severity of ulceration, a clinically important outcome.

Further analysis of the mucositis scores was performed by comparing the scores for the M40403 treated groups with the control on each day of scoring using the Mann-Whitney Rank Sum test. The results of this analysis are shown in Table 8. The groups treated with M40403 at 30 mg/kg/dose once daily from day −1 to day 3 (group 2), M40403 at 10 mg/kg/dose twice daily from day −1 to day 3 (group 4) and M40403 at 30 mg/kg/dose once daily from day 0 to day 3 (group 6) did not show any days with significant differences relative to controls. The scores for the group treated with M40403 at 3 mg/kg/dose twice daily from day −1 to day 3 (group 3) were significant different from controls on days 14 (P=0.013), 22 (P=0.047) and 28 (P=0.012). The group treated with M40403 at 30 mg/kg/dose twice daily from day −1 to day 3 (group 5) was significant different from controls on days 24 (P=0.048). The group treated with M40403 at 30 mg/kg/dose twice daily from day 0 to day 3 (group 7) had significantly lower scores than the control group on days 14 (P<0.001), 16 (P=0.001), 18 (P=0.007), 20 (P=0.001), 22 (P<0.001), 24 (P=0.006), 26 (P=0.001) and 28 (P=0.012). The group treated with M40403 at 30 mg/kg/dose twice daily on day 0 (group 8) had significantly lower scores than the control group on days 16 (P=0.001), 18 (P=0.033), 20 (P=0.002), 22 (P<0.001), 24 (P=0.010), 26 (P<0.001) and 28 (P=0.009). The group treated with M40403 at 30 mg/kg/dose twice daily on day 0 and on day 7 (group 9) had significantly lower scores than the control group on days 14 (P=0.004), 16 (P=0.047), 20 (P<0.001), 22 (P=0.015) and 26 (P=0.047).

TABLE 8 Day Group Comparison 6 8 10 12 14 16 18 20 22 24 26 28 Control v 0.467 0.655 0.944 0.499 0.097 0.231 0.233 0.315 0.235 0.294 0.273 0.253 M40403 ip 30 mg/kg qd day −1 to 3 Control v 0.985 0.231 0.985 0.748 0.013 0.297 0.865 0.459 0.047 0.610 0.283 0.012 M40403 ip 3 mg/kg bid day −1 to 3 Control v 0.230 0.985 0.985 0.865 0.506 0.955 0.437 0.414 0.354 0.595 0.463 0.584 M40403 ip 10 mg/kg bid day −1 to 3 Control v 0.725 0.655 0.410 0.428 0.097 0.141 0.216 0.097 0.170 0.048 0.053 0.066 M40403 ip 30 mg/kg bid day −1 to 3 Control v 0.961 0.944 0.798 0.834 0.361 0.870 0.592 0.981 0.437 0.388 0.362 0.798 M40403 ip 30 mg/kg qd day 0 to 3 Control v 0.776 0.555 0.472 0.940 <0.001  0.001 0.007 0.001 <0.001  0.006 0.001 0.012 M40403 ip 30 mg/kg bid day 0 to 3 Control v 0.776 0.985 0.374 0.519 0.061 0.001 0.033 0.002 <0.001  0.010 <0.001  0.009 M40403 ip 30 mg/kg bid day 0 Control v 0.415 0.721 0.156 0.851 0.004 0.047 0.091 <0.001  0.015 0.163 0.047 0.219 M40403 ip 30 mg/kg bid day 0 & 7 Table 4. ACT-03. The significance of group differences observed in daily mucositis scores was determined using the Mann-Whitney rank sum test. This nonparametric statistic is appropriate for the visual mucositis scoring scale. The p values for each calculation are shown. Significant improvements are shown underlined.

CONCLUSIONS

1. Based on observations of body weight, ip injections of M40403, appear to show some toxicity. The only groups that did not exhibit some statistically significant reductions in growth rate were those dosed with M40404 on day 0 only or on day 0 and day 7. Some mortality was noted on the day of radiation, but it is not clear whether this was related to treatment with M40403, or simply accidental death during the anesthesia process

2. The groups treated with M40403 at 30 mg/kg/dose once daily from day −1 to day 3, M40403 at 10 mg/kg/dose twice daily from day −1 to day 3 or M40403 at 30 mg/kg/dose once daily from day 0 to day 3 did not generally show any significant reduction in the course of mucositis relative to controls, although the group treated with M40403 at 30 mg/kg/dose once daily from day −1 to day 3 showed a significant reduction in the number of animals days with a score of 3 or higher (P=0.034).

3. The group treated with M40403 at 3 mg/kg/dose twice daily from day −1 to day 3 had significant lower mucositis scores from controls on days 14 (P=0.013), 22 (P=0.047) and 28 (P=0.012), and had a significant reduction in the number of animals days with a score of 3 or higher (P=0.020).

4. The group treated with M40403 at 30 mg/kg/dose twice daily from day −1 to day 3 had significantly lower scores than the control group on day 24 (P=0.048) and had a significant reduction in the number of animals days with a score of 3 or higher (P=0.007).

5. The group treated with M40403 at 30 mg/kg/dose twice daily from day 0 to day 3 (group 7) had significantly lower scores than the control group on days 14 (P<0.001), 16 (P=0.001), 18 (P=0.007), 20 (P=0.001), 22 (P<0.001), 24 (P=0.006), 26 (P=0.001) and 28 (P=0.012) and had a significant reduction in the number of animals days with a score of 3 or higher (P<0.001).

6. The group treated with M40403 at 30 mg/kg/dose twice daily on day 0 (group 8) had significantly lower scores than the control group on days 16 (P=0.001), 18 (P=0.033), 20 (P=0.002), 22 (P<0.001), 24 (P=0.010), 26 (P<0.001) and 28 (P=0.009) and had a significant reduction in the number of animals days with a score of 3 or higher (P<0.001).

7. The group treated with M40403 at 30 mg/kg/dose twice daily on day 0 and on day 7 had significantly lower scores than the control group on days 14 (P=0.004), 16 (P=0.047), 20 (P<0.001), 22 (P=0.015) and 26 (P=0.047) and had a significant reduction in the number of animals days with a score of 3 or higher (P<0.001).

Other Embodiments

The detailed description set-forth above is provided to aid those skilled in the art in practicing the present invention. However, the invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed because these embodiments are intended as illustration of several aspects of the invention. Any equivalent embodiments are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description which do not depart from the spirit or scope of the present inventive discovery. Such modifications are also intended to fall within the scope of the appended claims.

REFERENCES CITED

All publications, patents, patent applications and other references cited in this application are incorporated herein by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application or other reference was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Citation of a reference herein shall not be construed as an admission that such is prior art to the present invention.

-   1. Knox J J, Puodziunas A L, Feld R. Chemotherapy-induced oral     mucositis. Prevention and management. Drugs Aging 2000;     17(4):257-67. -   2. Peterson D E. Research advances in oral mucositis. Curr Opin     Oncol 1999; 11(4):261-6. -   3. Plevova P. Prevention and treatment of chemotherapy- and     radiotherapy-induced oral mucositis: a review. Oral Oncol 1999;     35(5):453-70. -   4. Sonis S T, Oster G, Fuchs H, Bellm L, Bradford W Z, Edelsberg J,     et al. Oral mucositis and the clinical and economic outcomes of     hematopoietic stem-cell transplantation. J Clin Oncol 2001;     19(8):2201-5. -   5. Eldor A, Fuks Z, Matzner Y, Witte L D, Vlodavsky I. Perturbation     of endothelial functions by ionizing irradiation: effects on     prostaglandins, chemoattractants and mitogens. Semin Thromb Hemost     1989; 15(2):215-25. -   6. Sonis S T, Van Vugt A G, McDonald J, Dotoli E, Schwertschlag U,     Szklut P, et al. Mitigating effects of interleukin 11 on consecutive     courses of 5-fluorouracil-induced ulcerative mucositis in hamsters.     Cytokine 1997; 9(8):605-12. -   7. Sonis S T, Van Vugt A G, Brien J P, Muska A D, Bruskin A M, Rose     A, et al. Transforming growth factor-beta 3 mediated modulation of     cell cycling and attenuation of 5-fluorouracil induced oral     mucositis. Oral Oncol 1997; 33(1):47-54. -   8. Sonis S T, Peterson R L, Edwards L J, Lucey C A, Wang L, Mason L,     et al. Defining mechanisms of action of interleukin-11 on the     progression of radiation-induced oral mucositis in hamsters. Oral     Oncol 2000; 36(4):373-81. -   9. Sonis S T, Tracey C, Shklar G, Jenson J, Florine D. An animal     model for mucositis induced by cancer chemotherapy. Oral Surg Oral     Med Oral Pathol 1990; 69(4):437-448

APPENDIX 1 Animal Weights DAY Group Animal −1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 Animal weights from a first study 1 1 69 92 92 90 101 102 107 110 113 110 118 119 121 110 120 1 2 94 101 101 108 109 109 116 116 118 122 121 125 126 123 134 1 3 93 96 96 99 102 105 107 104 112 115 115 118 120 118 118 1 4 68 92 90 93 96 98 99 101 102 106 106 109 110 132 114 1 5 98 98 98 102 106 109 113 108 117 123 123 127 129 130 130 1 6 97 102 101 107 109 111 116 118 121 123 125 127 130 131 130 1 7 87 95 90 94 97 98 105 104 101 105 105 105 107 110 109 1 8 91 96 97 100 102 103 105 109 111 115 115 117 119 126 126 2 9 100 dead 2 10 91 93 90 95 91 93 92 99 101 106 107 107 108 114 115 2 11 89 89 87 88 83 86 86 91 93 96 99 103 105 110 111 2 12 98 99 98 109 106 108 116 117 121 103 123 125 126 125 130 2 13 98 104 102 106 109 110 116 120 123 128 127 129 132 145 139 2 14 101 102 98 98 97 98 106 110 112 117 119 120 122 128 128 2 15 76 76 75 74 77 79 81 82 84 86 86 89 89 92 93 2 16 90 dead 3 17 93 99 98 98 98 103 105 103 106 108 107 111 113 116 118 3 18 94 95 96 97 98 100 103 107 108 109 107 106 108 115 116 3 19 96 96 100 101 105 107 109 118 114 115 113 116 119 121 121 3 20 99 99 102 97 108 105 113 117 120 124 123 125 127 134 134 3 21 91 93 95 104 99 102 105 107 110 115 113 113 116 120 123 3 22 105 104 95 110 112 115 120 124 128 132 130 138 138 144 145 3 23 92 93 93 97 99 102 103 104 106 107 110 112 116 114 117 3 24 93 91 93 90 98 99 101 105 107 109 107 107 110 101 105 4 25 96 97 95 93 98 98 102 106 107 110 111 113 115 119 118 4 26 89 89 88 90 93 92 97 99 101 104 103 102 103 110 108 4 27 93 98 93 98 102 106 106 107 109 116 118 119 120 123 119 4 28 99 100 99 98 100 103 106 110 112 115 118 121 122 122 125 4 29 90 91 90 96 100 102 100 101 104 107 107 108 110 112 111 4 30 93 98 95 99 102 103 106 111 111 116 116 119 121 125 122 4 31 93 94 89 91 95 98 99 103 106 108 109 112 116 116 114 4 32 91 92 91 95 98 100 102 102 105 109 111 112 114 114 116 5 33 74 dead 5 34 86 83 84 83 86 88 87 90 94 97 98 100 102 106 106 5 35 96 98 96 100 101 102 100 103 108 111 116 115 116 119 118 5 36 90 92 91 94 97 98 95 96 101 104 105 107 109 114 113 5 37 100 dead 5 38 96 101 100 98 101 103 104 108 110 114 117 119 120 128 129 5 39 87 84 82 79 78 79 81 84 86 91 93 94 99 97 98 5 40 89 82 79 78 79 82 81 88 87 91 94 98 100 101 102 6 41 94 92 98 97 99 102 98 98 103 108 109 110 112 116 115 6 42 94 95 93 98 99 101 104 103 109 111 113 114 116 119 119 6 43 101 101 101 107 109 110 115 117 122 126 127 129 130 134 134 6 44 90 dead 6 45 89 88 88 90 91 96 94 92 95 99 102 102 105 105 102 6 46 93 94 92 103 102 102 102 104 108 110 112 114 116 119 119 6 47 99 101 98 97 108 109 111 111 119 118 119 123 126 125 126 6 48 91 dead 7 49 91 94 91 95 101 104 98 102 106 107 108 113 115 119 120 7 50 93 95 92 91 93 93 98 104 103 105 106 109 111 115 116 7 51 90 87 86 87 97 98 95 96 99 101 103 107 108 113 113 7 52 95 99 95 99 102 104 107 108 113 115 114 113 115 120 120 7 53 92 93 88 89 91 93 90 93 97 99 101 102 106 109 108 7 54 98 101 98 102 105 108 101 108 109 112 113 116 119 121 122 7 55 91 93 90 93 99 101 98 101 104 108 109 111 113 117 118 7 56 88 87 81 83 78 81 78 80 82 85 86 92 95 94 99 8 57 94 97 93 99 102 105 105 108 112 113 114 117 119 120 121 8 58 88 90 84 89 93 95 98 104 105 106 107 111 114 116 118 8 59 88 92 84 91 93 97 97 100 102 105 103 105 108 110 112 8 60 86 89 86 96 95 96 99 100 105 106 106 108 111 114 114 8 61 94 94 91 97 99 103 103 106 110 113 116 117 119 125 124 8 62 93 94 88 93 98 101 104 108 111 108 112 118 120 122 121 8 63 91 94 91 96 99 100 103 106 109 110 113 115 117 118 120 8 64 98 101 96 101 107 107 111 113 117 118 119 125 127 124 124 9 65 98 101 100 104 108 110 113 115 116 114 117 119 131 124 124 9 66 94 98 94 98 102 105 107 110 112 113 114 117 118 125 124 9 67 89 93 90 95 98 101 104 106 108 105 109 112 115 115 116 9 68 93 96 91 95 100 103 105 104 111 109 112 113 116 117 116 9 69 98 99 98 102 106 109 110 114 118 119 121 124 129 131 129 9 70 92 dead 9 71 91 91 89 94 96 100 102 105 106 104 107 110 111 116 120 9 72 95 96 95 98 101 105 106 109 111 111 114 114 116 116 120 Animal weights from a second study 1 1 88 92 92 94 96 98 102 103 103 106 108 110 114 113 116 1 2 89 93 92 96 100 102 103 107 109 110 114 116 119 117 120 1 3 96 98 98 101 105 108 110 112 115 120 122 125 126 125 123 1 4 95 97 97 102 103 107 108 111 114 117 118 121 125 123 128 1 5 88 91 88 91 94 97 98 100 103 105 106 110 112 109 111 1 6 90 95 93 98 101 104 107 108 111 114 115 122 122 121 124 1 7 88 89 88 91 94 96 92 99 103 102 104 108 110 108 111 1 8 92 AOD 2 9 85 86 86 90 93 96 95 110 104 107 109 111 113 112 113 2 10 66 89 86 91 94 96 97 100 104 106 107 108 112 112 114 2 11 68 91 91 94 96 97 98 103 104 105 107 109 112 112 115 2 12 100 104 102 106 105 112 113 116 121 121 124 129 132 129 132 2 13 77 81 80 84 87 83 94 95 97 98 101 104 107 108 108 2 14 92 98 96 101 103 106 108 112 111 115 116 117 118 117 120 2 15 68 91 69 93 95 95 97 101 102 104 106 109 111 113 115 2 16 94 97 97 100 102 105 106 113 112 115 116 120 123 123 126 3 17 90 99 90 96 97 96 96 90 99 90 97 101 100 90 99 3 18 68 91 86 69 92 94 94 94 98 99 96 97 93 91 93 3 19 90 93 90 90 92 94 82 80 84 84 85 82 87 dead 3 20 68 91 85 69 82 84 88 79 81 80 76 80 81 80 62 3 21 101 102 98 100 101 103 101 101 107 108 104 107 108 103 106 3 22 92 96 87 93 92 95 94 88 91 86 82 83 80 78 80 3 23 65 dead 3 24 92 95 91 95 97 99 99 100 104 103 100 101 100 98 100 4 25 86 90 88 91 93 95 99 97 102 105 108 109 105 110 114 4 26 92 95 93 99 102 103 106 109 112 113 116 121 120 119 123 4 27 85 87 86 89 90 94 94 98 99 102 119 87 107 105 108 4 28 94 96 96 101 103 106 107 111 113 115 119 120 127 125 127 4 29 94 96 96 99 101 104 107 110 112 117 117 123 122 121 123 4 30 91 94 95 99 102 104 104 100 109 113 115 117 119 118 119 4 31 93 85 84 38 89 93 94 99 97 98 100 105 105 104 106 4 32 74 74 75 79 79 82 86 87 92 94 95 103 98 95 98 5 33 83 83 83 89 89 98 95 98 101 104 105 115 110 108 111 5 34 79 80 79 83 86 88 90 93 95 102 99 101 104 103 106 5 35 83 87 85 90 93 95 97 94 101 102 103 105 106 105 109 5 36 84 86 86 91 93 97 100 102 107 111 110 109 115 118 114 5 37 85 86 85 38 89 91 93 102 99 101 102 106 108 107 111 5 38 78 79 79 32 84 88 88 97 95 101 99 103 105 102 104 5 39 85 87 87 90 94 94 99 101 104 106 107 111 112 111 113 5 40 81 83 82 37 89 90 93 95 97 100 101 107 107 107 109 6 41 86 87 81 35 93 97 78 82 85 89 91 96 98 100 102 6 42 91 90 88 93 95 99 102 107 112 114 113 120 122 120 123 6 43 84 87 84 33 79 79 84 93 89 93 95 98 102 103 107 6 44 91 94 92 92 93 94 96 101 104 106 103 113 114 114 119 6 45 78 dead 6 46 86 89 dead 6 47 89 93 86 91 79 91 92 99 101 105 109 112 114 114 116 6 48 88 92 87 89 92 86 90 97 100 109 105 111 114 114 118 DAY Group Animal 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 Animal weights from a first study 1 1 120 120 130 132 133 137 138 139 143 140 144 140 149 100 148 1 2 134 134 138 140 144 148 151 151 168 156 158 160 162 162 164 1 3 119 123 124 127 130 131 129 133 135 137 139 142 144 142 138 1 4 115 118 117 121 124 126 134 129 132 133 132 133 138 138 139 1 5 133 135 138 141 142 148 147 150 153 154 158 165 163 163 159 1 6 131 131 136 138 141 141 142 143 146 149 149 152 153 171 157 1 7 111 112 116 116 120 121 122 123 127 128 128 130 134 128 138 1 8 129 130 130 132 137 137 140 141 145 147 148 150 153 154 154 2 9 2 10 116 116 119 121 121 122 124 125 127 129 132 137 133 135 135 2 11 112 113 118 120 121 124 128 127 133 131 138 160 140 142 143 2 12 131 133 135 133 142 147 149 151 165 156 158 134 165 164 168 2 13 140 142 144 147 149 154 150 156 160 162 185 166 186 168 174 2 14 129 129 132 135 125 138 139 140 144 145 148 150 151 152 155 2 15 99 95 99 101 101 103 108 106 109 111 109 110 114 115 116 2 16 3 17 117 118 119 121 124 124 126 127 129 130 132 133 136 138 139 3 18 116 118 120 123 125 128 130 132 136 138 140 142 145 149 148 3 19 122 125 128 131 132 135 137 138 140 143 143 140 147 149 151 3 20 132 134 138 140 143 145 148 149 152 154 156 160 160 162 165 3 21 123 125 127 129 132 132 134 136 138 142 143 144 148 147 148 3 22 149 150 155 158 160 164 166 169 172 174 176 179 181 183 182 3 23 117 118 117 120 120 126 126 128 130 131 132 136 136 139 140 3 24 105 110 112 115 117 118 124 126 129 132 134 137 138 138 143 4 25 121 124 128 130 131 132 133 134 137 139 140 142 145 145 144 4 26 109 110 112 117 111 117 119 119 121 128 125 126 127 130 128 4 27 123 124 128 131 130 135 138 137 143 146 145 146 150 156 152 4 28 128 129 133 135 137 139 140 140 144 145 148 150 152 151 152 4 29 117 119 121 124 125 129 131 128 133 134 136 140 141 140 142 4 30 121 126 131 134 134 136 141 141 145 148 149 152 154 158 157 4 31 117 120 123 125 128 127 131 130 132 132 134 136 138 130 139 4 32 118 120 125 126 127 130 133 134 137 138 141 143 145 133 148 5 33 5 34 107 108 109 111 120 118 120 120 126 129 128 130 134 134 134 5 35 120 119 123 125 129 129 130 129 134 137 137 139 140 141 142 5 36 116 117 119 121 120 125 127 127 131 133 133 135 139 134 140 5 37 5 38 134 134 138 140 152 144 148 144 149 152 154 156 159 161 161 5 39 100 103 106 108 117 113 116 115 121 122 123 124 130 132 130 5 40 104 103 104 105 116 113 112 111 115 117 118 119 125 128 124 6 41 114 116 117 119 129 125 127 128 136 134 136 140 142 151 144 6 42 120 121 124 127 133 132 133 133 141 140 142 143 146 149 150 6 43 135 138 141 142 150 150 154 155 160 160 164 165 172 172 172 6 44 6 45 101 98 100 103 105 104 108 106 111 111 111 113 115 114 115 6 46 122 123 126 128 132 131 134 132 138 139 141 145 146 159 144 6 47 130 131 135 137 138 140 143 142 147 150 152 153 156 141 160 6 48 7 49 127 130 131 133 135 140 141 142 148 149 150 152 156 156 156 7 50 118 117 124 127 120 128 129 129 135 137 138 140 143 142 146 7 51 118 121 118 121 125 127 133 131 137 138 138 140 144 146 145 7 52 124 122 128 131 132 133 137 136 143 142 143 145 148 140 149 7 53 110 112 117 120 118 121 124 124 128 129 130 133 135 138 137 7 54 125 126 129 129 135 137 138 138 142 144 146 147 151 145 152 7 55 120 123 125 127 130 130 132 133 139 140 142 145 148 146 150 7 56 100 102 105 108 111 112 113 113 116 121 123 124 127 121 130 8 57 123 125 128 131 138 132 137 137 139 140 143 143 147 147 148 8 58 119 120 120 124 128 128 131 131 134 134 135 137 139 135 142 8 59 111 115 118 121 122 118 122 121 123 126 127 131 129 126 127 8 60 117 118 122 124 125 128 131 131 136 138 139 143 145 146 146 8 61 126 128 126 130 138 137 139 141 142 144 144 146 151 151 153 8 62 122 126 128 130 134 133 137 138 138 142 145 147 147 154 153 8 63 123 125 127 131 129 132 135 135 137 138 139 147 143 145 146 8 64 125 126 127 130 131 135 137 135 139 142 142 144 147 142 151 9 65 127 126 133 137 135 136 138 139 141 142 144 146 149 148 151 9 66 127 129 130 134 137 139 141 139 143 145 148 150 152 144 152 9 67 119 118 123 126 125 126 130 130 132 135 136 137 140 128 143 9 68 120 119 121 124 129 127 130 129 131 122 133 136 134 122 136 9 69 133 134 138 141 142 146 152 149 151 153 155 159 162 165 166 9 70 9 71 122 122 125 128 128 131 143 134 137 139 139 141 145 142 147 9 72 120 120 121 124 133 128 129 125 129 132 154 137 138 138 140 Animal weights from a second study 1 1 115 115 117 119 119 114 115 116 117 118 121 122 123 124 127 1 2 124 124 125 129 131 131 135 136 136 138 140 141 147 149 148 1 3 131 132 137 142 145 144 148 152 153 155 159 161 161 163 169 1 4 124 124 124 129 130 131 134 138 139 140 143 147 145 149 152 1 5 111 112 113 117 119 119 123 127 127 128 131 132 133 136 136 1 6 127 125 124 128 130 130 134 136 139 141 128 147 148 149 153 1 7 113 112 114 116 117 118 123 123 124 125 145 128 139 133 134 1 8 2 9 116 115 121 123 125 127 133 135 138 139 142 144 146 149 152 2 10 118 117 118 121 123 125 129 130 132 135 139 142 143 147 148 2 11 119 116 117 119 120 121 125 126 128 129 131 134 134 138 139 2 12 134 135 135 138 142 140 145 144 150 150 154 155 158 162 160 2 13 112 111 113 117 119 117 122 125 124 122 129 130 132 136 136 2 14 118 120 122 123 127 128 131 134 135 136 140 144 142 144 150 2 15 118 120 121 123 124 126 129 131 132 135 135 137 138 140 142 2 16 127 126 128 129 131 131 135 136 136 140 143 145 143 113 151 3 17 99 97 94 90 102 102 100 110 115 117 121 124 124 127 131 3 18 91 86 84 84 86 91 95 99 101 104 108 111 112 115 118 3 19 3 20 83 78 77 79 81 85 89 93 97 99 103 99 108 110 115 3 21 103 99 99 98 106 107 112 115 119 117 127 128 129 132 135 3 22 79 74 72 75 79 81 85 88 91 93 97 106 107 108 109 3 23 3 24 102 99 101 105 108 110 114 117 121 123 127 130 132 135 138 4 25 115 116 119 124 125 123 129 131 133 133 137 138 140 145 146 4 26 123 124 129 134 134 133 141 144 145 143 150 151 152 156 160 4 27 110 110 111 114 116 114 118 121 123 124 126 130 131 133 136 4 28 129 127 129 132 132 135 138 142 143 143 128 148 158 148 154 4 29 125 125 130 132 135 135 142 142 145 149 151 153 157 160 161 4 30 122 120 126 127 130 127 134 136 139 139 142 142 144 146 149 4 31 111 109 112 114 116 117 120 123 124 122 129 130 130 133 135 4 32 107 104 108 111 112 112 116 119 122 126 125 125 124 126 132 5 33 112 111 114 118 119 120 124 124 124 128 131 132 138 141 140 5 34 111 110 114 118 119 121 123 127 126 128 132 133 134 137 139 5 35 120 113 114 116 116 119 124 126 125 128 130 132 137 134 138 5 36 120 121 124 127 130 132 135 137 139 144 145 148 150 152 158 5 37 110 108 111 115 117 116 121 123 124 124 129 129 131 134 136 5 38 107 108 110 114 117 116 122 125 125 128 131 134 133 137 139 5 39 113 118 121 125 126 127 130 132 132 134 136 137 146 142 148 5 40 112 110 113 116 116 118 121 123 124 125 128 128 130 133 135 6 41 108 107 110 112 116 117 120 121 123 125 128 128 130 136 136 6 42 118 117 115 108 106 104 104 103 134 104 106 108 108 109 110 6 43 109 108 112 115 115 116 122 124 124 125 127 130 129 131 134 6 44 120 120 123 125 127 128 132 133 135 137 137 138 136 139 142 6 45 6 46 6 47 124 124 127 129 133 133 137 137 136 138 141 143 142 150 146 6 48 122 122 126 129 129 131 135 137 138 139 142 145 143 146 149

APPENDIX 2 Mucositis Scores Cheek pouch photographs were scored by 2 independent scorers in a blinded manner resulting in 2 scores for each animal at each time point. DAY Group Animal 6 8 10 12 14 16 18 20 22 24 26 28 ACT-03 Blinded Scores Mucositis scores from a first study 1 1 0 0 0 2 3 3 3 3 3 2 2 1 1 1 0 0 0 1 3 3 3 3 3 2 1 1 1 2 1 0 1 2 3 3 4 3 3 3 2 2 1 2 0 0 0 2 3 3 3 3 3 3 2 1 1 3 1 0 1 2 3 4 3 3 3 2 2 2 1 3 0 0 0 1 3 4 3 3 3 1 2 1 1 4 1 0 0 2 3 3 3 4 2 2 2 2 1 4 0 0 0 1 3 3 3 3 2 2 2 1 1 5 0 1 0 2 3 3 3 3 3 2 2 2 1 5 0 0 1 2 3 3 3 3 3 1 1 1 1 6 0 0 1 2 3 3 4 3 2 2 2 2 1 6 0 0 1 2 3 3 3 3 2 2 2 1 1 7 0 0 1 2 3 3 2 2 1 1 1 1 1 7 0 0 1 2 3 3 1 1 1 1 1 1 1 8 1 0 1 1 2 3 3 3 3 2 2 2 1 8 0 0 1 0 2 3 3 3 3 2 2 1 2 9 dead 2 9 2 10 0 0 1 2 2 3 2 2 1 1 1 1 2 10 0 0 1 1 2 3 2 2 1 1 1 1 2 11 0 0 1 2 3 3 3 3 3 2 1 1 2 11 0 0 0 1 3 3 3 3 3 2 2 2 2 12 0 0 1 1 3 3 3 3 2 2 2 1 2 12 0 0 0 1 3 3 3 3 2 1 2 1 2 13 0 0 1 2 3 3 3 3 3 2 2 1 2 13 0 1 0 2 3 3 3 3 3 2 2 1 2 14 0 0 1 2 2 3 3 3 2 2 1 0 2 14 1 1 0 1 2 3 3 3 2 2 2 1 2 15 0 0 1 2 2 2 2 2 2 1 1 1 2 15 0 0 0 1 2 2 2 2 2 1 1 2 2 16 dead 2 16 3 17 0 0 1 1 1 3 3 3 2 2 1 0 3 17 1 0 1 1 2 3 3 3 2 2 2 1 3 18 0 0 1 1 2 3 3 3 3 3 2 0 3 18 1 1 1 1 2 3 3 3 3 3 2 1 3 19 0 0 1 2 3 3 3 3 2 2 2 0 3 19 1 1 0 2 3 3 3 3 2 2 1 2 3 20 0 0 1 1 2 3 4 3 2 1 2 1 3 20 0 0 1 2 2 3 3 3 2 1 1 2 3 21 0 0 0 1 2 2 2 2 2 1 2 0 3 21 0 0 0 1 2 2 2 2 2 2 2 1 3 22 0 0 1 2 1 2 2 2 1 1 1 0 3 22 0 1 0 1 2 2 2 2 2 2 1 1 3 23 0 0 0 3 3 4 4 3 2 2 2 0 3 23 0 1 0 3 3 4 4 3 2 2 1 1 3 24 0 0 1 2 3 3 3 3 2 1 2 0 3 24 1 1 0 2 3 3 3 3 2 1 1 1 4 25 0 0 1 2 1 1 1 1 1 1 2 1 4 25 0 0 0 2 1 1 1 1 2 2 1 2 4 26 0 0 0 0 3 3 3 3 2 1 2 1 4 26 0 0 0 0 3 3 3 3 2 2 1 2 4 27 0 0 1 2 2 3 3 3 3 2 2 0 4 27 0 1 0 2 2 4 3 3 3 2 2 1 4 28 0 0 2 2 3 3 3 3 3 2 2 1 4 28 0 0 2 2 3 3 3 3 3 2 2 2 4 29 0 0 1 2 3 3 4 3 2 2 2 0 4 29 0 0 0 1 3 4 3 3 2 2 1 1 4 30 0 0 1 2 3 3 3 3 2 2 2 0 4 30 0 0 0 1 3 4 3 3 2 2 2 2 4 31 0 0 1 2 3 3 2 2 2 2 2 1 4 31 0 0 0 2 3 3 2 2 2 2 1 2 4 32 0 0 1 2 3 3 3 3 3 2 2 1 4 32 0 0 0 2 3 4 3 3 3 2 1 2 5 33 dead 5 33 5 34 0 0 0 2 3 3 3 2 2 1 1 0 5 34 0 0 0 2 3 3 3 2 2 2 1 1 5 35 0 0 1 2 3 3 3 3 2 2 2 1 5 35 0 0 0 2 3 4 3 3 2 2 1 2 5 36 0 0 1 2 2 2 2 2 0 1 1 1 5 36 0 0 0 1 2 2 2 2 1 1 0 2 5 37 dead 5 37 5 38 0 0 0 2 2 2 2 2 2 0 2 0 5 38 1 0 0 1 2 2 1 2 1 0 1 1 5 39 0 0 0 1 3 3 3 3 3 1 1 0 5 39 0 1 0 0 3 3 3 3 3 0 1 1 5 40 1 0 2 1 2 3 3 3 3 2 2 0 5 40 0 1 1 1 2 3 3 3 3 2 2 1 6 41 0 0 1 1 1 3 3 3 3 2 2 0 6 41 1 0 1 0 3 3 3 3 3 2 2 1 6 42 0 0 1 2 3 3 3 4 2 2 2 1 6 42 0 0 1 1 3 3 3 3 2 2 1 2 6 43 0 0 0 2 3 3 2 2 2 2 2 1 6 43 1 0 0 2 3 3 2 2 2 1 2 2 6 44 dead 6 44 6 45 0 0 1 2 2 4 4 4 4 4 3 2 6 45 1 0 0 2 2 4 4 4 4 4 3 2 6 46 0 0 0 2 3 3 3 3 2 2 2 0 6 46 0 0 0 2 3 3 3 3 2 2 2 1 6 47 0 0 1 2 2 3 2 2 0 2 2 2 6 47 0 1 0 0 2 3 2 2 1 2 1 1 6 48 dead 6 48 7 49 0 0 1 2 2 1 1 1 0 1 1 1 7 49 0 1 0 2 2 1 0 0 1 0 1 1 7 50 0 0 0 2 2 2 3 1 1 0 0 0 7 50 1 0 0 2 2 1 3 0 2 1 1 1 7 51 0 0 1 1 2 3 1 2 1 0 0 0 7 51 1 0 0 0 2 3 2 1 2 0 1 1 7 52 0 0 0 2 2 2 2 2 1 0 0 0 7 52 1 1 0 2 2 2 1 0 2 1 1 1 7 53 0 0 1 1 2 3 3 3 2 1 1 0 7 53 0 1 0 0 2 3 3 3 2 2 1 1 7 54 0 0 2 2 2 3 3 3 2 0 1 0 7 54 0 0 0 2 2 3 3 3 1 2 1 2 7 55 0 0 0 2 3 2 2 2 2 1 2 0 7 55 0 0 0 2 3 2 2 1 2 2 1 2 7 56 0 0 1 2 2 2 1 1 1 2 2 0 7 56 0 0 1 1 2 2 1 1 1 2 1 1 8 57 0 0 0 2 3 3 3 3 1 1 0 1 8 57 0 1 0 2 3 3 3 3 1 2 0 2 8 58 1 0 0 2 2 2 2 2 1 1 1 1 8 58 1 0 0 2 2 2 2 2 1 2 0 1 8 59 0 0 1 2 1 1 1 1 1 1 2 0 8 59 1 0 0 2 1 2 1 2 1 1 2 1 8 60 0 0 1 1 3 3 3 1 1 0 0 0 8 60 0 0 0 1 3 3 3 2 1 2 0 1 8 61 0 0 1 2 2 2 3 2 1 1 1 0 8 61 0 0 0 2 2 2 3 1 1 1 1 1 8 62 0 0 0 2 3 2 1 2 2 0 1 0 8 62 0 0 0 1 3 2 2 1 2 2 0 1 8 63 0 0 1 2 2 2 1 2 2 0 1 0 8 63 1 0 1 2 2 2 1 1 2 2 0 1 8 64 0 0 1 2 3 3 3 3 2 0 2 0 8 64 1 0 0 2 3 3 3 3 2 1 0 1 9 65 0 0 1 2 2 3 3 2 2 0 2 1 9 65 1 0 0 2 2 3 3 2 2 1 1 2 9 66 0 0 0 2 2 2 2 2 0 1 1 0 9 66 1 0 0 1 1 1 1 1 1 2 0 1 9 67 0 0 0 2 1 1 2 1 0 0 1 0 9 67 1 0 0 1 1 1 1 1 1 1 0 1 9 68 1 1 2 2 2 3 3 2 1 1 2 0 9 68 1 0 0 1 2 3 3 1 2 2 1 1 9 69 0 0 0 2 3 3 3 3 3 3 2 1 9 69 1 1 0 1 3 3 3 3 3 3 1 2 9 70 9 70 9 71 0 0 1 2 2 2 2 2 2 1 2 0 9 71 0 0 0 2 2 2 2 2 2 2 1 2 9 72 0 0 0 2 3 4 3 1 2 2 2 1 9 72 0 0 0 2 3 3 3 2 2 1 1 2 ACT-03 Blinded Scores Mucositis scores from a second study 1 1 0 1 1 2 2 2 2 2 2 1 0 0 1 1 0 1 1 2 2 2 2 2 2 1 0 0 1 2 0 1 1 2 3 3 2 2 2 2 2 1 1 2 0 1 1 2 3 3 2 2 2 2 2 1 1 3 0 1 1 2 3 3 3 3 3 2 2 2 1 3 0 1 1 2 3 3 3 3 3 2 2 2 1 4 0 1 1 2 3 3 3 3 3 2 2 1 1 4 0 1 1 2 3 3 3 3 3 2 2 1 1 5 0 1 2 2 3 3 3 3 3 2 2 1 1 5 0 1 2 2 3 3 3 3 3 2 2 1 1 6 0 0 1 2 3 3 3 3 3 3 2 2 1 6 0 0 1 2 3 3 3 3 3 3 2 2 1 7 1 1 1 1 2 3 3 3 2 2 2 1 1 7 1 1 1 1 2 3 3 3 2 2 2 1 1 8 1 8 2 9 0 1 1 2 3 3 3 3 2 2 2 2 2 9 0 1 1 2 3 3 3 3 2 2 2 2 2 10 1 0 1 2 3 3 3 3 3 2 2 2 2 10 1 0 1 2 3 3 3 3 3 2 2 2 2 11 0 1 1 1 3 3 3 3 3 3 2 2 2 11 0 1 1 1 3 3 3 3 3 3 2 2 2 12 1 1 1 2 3 3 3 3 3 3 2 1 2 12 1 1 1 2 3 3 3 3 3 3 2 1 2 13 1 1 1 1 3 3 3 3 2 2 3 2 2 13 0 1 1 1 3 3 3 3 2 2 3 2 2 14 0 1 2 2 3 3 3 3 2 2 2 1 2 14 0 1 2 2 3 3 3 3 2 2 2 1 2 15 1 1 1 1 1 2 2 1 2 2 2 2 2 15 0 1 1 1 1 2 2 1 2 2 2 2 2 16 1 0 1 2 2 2 3 3 3 2 1 1 2 16 1 0 1 2 2 2 3 3 3 2 1 1 3 17 0 1 1 2 2 3 3 3 3 3 2 2 3 17 0 1 1 2 2 3 3 3 3 3 2 2 3 18 1 1 1 2 3 3 3 3 3 3 1 1 3 18 1 1 1 2 3 3 3 3 3 3 1 1 3 19 1 1 1 1 3 19 1 1 1 1 3 20 1 1 1 1 3 3 3 3 3 3 3 2 3 20 1 1 1 1 3 3 3 3 3 3 3 2 3 21 0 1 1 1 3 3 3 3 3 3 2 2 3 21 0 1 1 1 3 3 3 3 3 3 2 2 3 22 1 1 2 2 3 3 3 3 2 2 2 1 3 22 1 1 2 2 3 3 3 3 2 2 2 1 3 23 3 23 3 24 0 0 1 1 2 2 3 2 2 1 1 1 3 24 0 0 1 1 2 2 3 2 2 1 1 1 

What is claimed is:
 1. A method comprising parenterally administering a dose of a pentaaza macrocyclic ring complex to a mammal to lessen the severity of a mucositis affecting the mucosal lining of the gastrointestinal tract, wherein the mammal is afflicted with cancer and the dose of the pentaaza macrocyclic ring complex is administered on the day before or the day of, but prior to or simultaneous with the mammal receiving either or both of (i) a dose of chemotherapy treatment, or (ii) a dose of radiation therapy, the dose of the pentaaza macrocyclic ring complex being therapeutically sufficient, without requiring the administration of any additional doses of the pentaaza macrocyclic ring complex after either or both of the dose of chemotherapy treatment or dose of radiation therapy, to lessen the severity of the mucositis affecting the mucosal lining of the gastrointestinal tract resulting from the delivered chemotherapy and/or radiation dose, and wherein the pentaaza macrocyclic ring complex is a compound represented by the formula:

wherein W is an unsubstituted pyridino moiety; U and V are trans-cyclohexanyl fused rings; R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀ are each independently selected from the group consisting of hydrogen, alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and heterocyclyl, and aralkyl radicals and radicals attached to the α-carbon or α-amino acids, M is Mn²⁺ or Mn³⁺; and X, Y and Z are independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino, heterocycloaryl amino, amine oxides, hydrazine, alkyl hydrazine, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid, aryl carboxylic acid, urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate, thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkylaryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkylaryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalomanganate, tetrafluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion exchange resins, or the corresponding anions thereof; or X, Y and Z are independently selected from the group consisting of charge-neutralizing anions which are derived from any monodentate or polydentate coordinating ligand and a ligand system and the corresponding anion thereof; or X, Y and Z are independently attached to one or more of R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀; and n is an integer from 0 to
 3. 2. A method in accordance with claim 1, wherein M is Mn²⁺.
 3. A method in accordance with claim 1, wherein R₁, R₂, R′₂, R₃, R₄, R₅, R′₅, R₆, R′₆, R₇, R₈, R₉, R′₉, and R₁₀ are each hydrogen.
 4. A method in accordance with claim 1, wherein the mammal is a human patient.
 5. A method in accordance with claim 1, wherein the pentaaza macrocyclic ring complex is administered prior to or simultaneous with the dose of chemotherapy.
 6. A method in accordance with claim 1, wherein the pentaaza macrocyclic ring complex is administered to the mammal prior to the mammal receiving the dose of chemotherapy.
 7. A method in accordance with claim 5, wherein the pentaaza macrocyclic ring complex is administered to the mammal simultaneously with the mammal receiving the dose of chemotherapy.
 8. A method in accordance with claim 1, wherein the pentaaza macrocyclic ring complex is administered prior to or simultaneous with the dose of radiation therapy.
 9. A method in accordance with claim 8, wherein the pentaaza macrocyclic ring complex is administered to the mammal prior to the mammal receiving a dose of radiation therapy.
 10. A method in accordance with claim 8, wherein the pentaaza macrocyclic ring complex is administered to the mammal simultaneously with the mammal receiving the dose of radiation therapy.
 11. A method in accordance with claim 1, wherein an initial dose of the pentaaza macrocyclic ring complex is administered prior to onset of injury to the mucosal lining resulting from the delivered chemotherapy and/or radiation dose.
 12. A method in accordance with claim 1, wherein the pentaaza macrocyclic ring complex is administered intravenously or subcutaneously.
 13. A method in accordance with claim 12, wherein the pentaaza macrocyclic ring complex is administered intravenously.
 14. A method in accordance with claim 1, wherein the mammal is afflicted with a tumor of the head and/or neck. 